Methods and compositions for treating viral or virally-induced conditions

ABSTRACT

Provided are methods and compositions for the prevention and/or treatment of viral conditions, virally-induced conditions and inflammatory conditions. The methods can comprise administering to a subject a viral inducing agent with an antiviral agent, and optionally an additional agent. The viral inducing agent can be a HDAC inhibitor administered orally.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation application of co-pending U.S. patentapplication Ser. No. 15/335,776 filed Oct. 27, 2016, which is acontinuation application of U.S. patent application Ser. No. 14/728,592filed Jun. 2, 2015, which is a continuation application of U.S. patentapplication Ser. No. 13/912,637 filed Jun. 7, 2013, which is acontinuation application of U.S. patent application Ser. No. 13/046,555filed on Mar. 11, 2011, which claims the benefit under 35 U.S.C. §119(e) of U.S. Provisional Application No. 61/430,931 filed on Jan. 7,2011, and U.S. Provisional Application No. 61/313,052 filed on Mar. 11,2010, the contents of each of which are incorporated herein by referencein their entireties.

In compliance with 37 C.F.R. § 1.71(g)(1), disclosure is herein madethat the claimed invention was made pursuant to a Joint ResearchAgreement as defined in 35 U.S.C. 103(c)(3), that was in effect on orbefore the date the claimed invention was made, and as a result ofactivities undertaken within the scope of the Joint Research Agreement,by or on the behalf of Boston University and Hemaquest Pharmaceuticals,Inc., now Viracta Therapeutics, Inc.

SEQUENCE LISTING

The instant application contains a Sequence Listing which has beensubmitted in ASCII format via EFS-Web and is hereby incorporated byreference in its entirety. Said ASCII copy, created on Apr. 19, 2018, isnamed 701586-070085_SL.txt and is 1,296 bytes in size.

BACKGROUND OF THE INVENTION

Many patients can have latent infections in which a virus is present butis not expressing viral proteins such as viral thymidine kinase orprotein kinase, the target for common anti-viral drugs such as acyclovirand ganciclovir. A viral inducing drug such as a histone deacetylaseinhibitor (HDAC inhibitor—HDACi) can be used to re-induce the expressionof viral thymidine kinase or protein kinase in viral infected cells inthe subject; the subject can then be treatment with antiviral drugs toeliminate latent viral infections. As EBV and/or other latent viralinfections can be associated with a variety of conditions, many of whichare inflammatory conditions, such as lymphomas, autoimmune conditions,allergic conditions, eliminating the latent virus with this therapy canbe used to prevent or treat such conditions.

EBV can induce autoimmune conditions through a number of potentialmechanisms, e.g.: 1) activating B cells to produce auto-antibodies, 2)turning on T cells that attack host tissue, 3) molecular mimicry inwhich EBV antigens cross react with host antigens such that autoimmunecondition results when T cells or antibodies reactive with theseantigens cross react with host antigens causing damage to host tissues,4) EBV infected B cells produce cytokines which turn on other elementsof the immune system and increase inflammation which can also exacerbateautoimmune condition, 5) EBV infected B or T cells can becomeimmortalized via EBV proteins indirectly turning out anti-apoptotic orsurvival pathways. Autoimmune conditions with evidence of an EBVrelationship include multiple sclerosis, systemic lupus erythematosus,rheumatoid arthritis and Sjogren's syndrome. Cytomegalovirus and herpessimplex virus have been associated with coronary artery condition.

There is a need for methods of treating and/or preventing viralconditions, viral-induced conditions, and related inflammatoryconditions.

SUMMARY OF THE INVENTION

In one aspect, a method for treating and/or preventing a viralcondition, a virally-induced condition, or an inflammatory conditioncomprising administering a histone deacetylase inhibitor (HDACinhibitor) and an antiviral agent wherein the HDAC inhibitor is apyrimidine hydroxamic acid derivative is provided.

In another aspect, a method for treating and/or preventing a viral orvirally-induced condition is provided comprising administering a HDACinhibitor and an antiviral agent wherein the viral or virally-inducedcondition is caused by a DNA virus and the HDAC inhibitor isadministered at dose of less than 2 mg/kg per dose.

In yet another aspect, a method for treating and/or preventing a viralcondition, a virally-induced condition, or an inflammatory condition isprovided comprising administering a HDAC inhibitor and an antiviralagent wherein the MW of the HDAC inhibitor is greater than 275 g/mol.

In another aspect, a method for treating and/or preventing a viralcondition or a virally-induced condition is provided, comprisingadministering a HDAC inhibitor wherein the HDAC inhibitor is apyrimidine hydroxamic acid derivative.

In another aspect, a composition comprising a (i) HDAC inhibitor and(ii) an antiviral agent is provided, wherein the HDAC inhibitor is apyrimidine hydroxamic acid derivative.

Provided herein, in one aspect, is a method for treating and/orpreventing a viral or virally-induced condition comprising administeringa HDAC inhibitor and an antiviral agent wherein the viral orvirally-induced condition is caused by a DNA virus and the HDACinhibitor is administered at dose of less than 2 mg/kg per dose. In someembodiments, the HDAC inhibitor is Vorinostat/suberoyl anilidehydroxamic acid, JNJ-26481585(N-hydroxy-2-(4-((((1-methyl-1H-indol-3-yl)methyl)amino)methyl)piperidin-1-yl)pyrimidine-5-carboxamide),R306465/JNJ-16241199(N-hydroxy-5-(4-(naphthalen-2-ylsulfonyl)piperazin-1-yl)pyrimidine-2-carboxamide), CHR-3996(2-(6-{[(6-Fluoroquinolin-2-yl)methyl]amino}-3-azabicyclo[3.1.0]hex-3-yl)-N-hydroxypyrimidine-5-carboxamide),Belinostat/PXD101, Panobinostat/LBH-589, trichostatin A/TSA(7-[4-(dimethylamino)phenyl]-N-hydroxy-4,6-dimethyl-7-oxohepta-2,4-dienamide),ITF2357, CBHA, Givinostat/ITF2357, romidepsin (Istodax™), PCI-24781,depsipeptide (FR901228 or FK228), butyrate, phenylbutyrate, valproicacid, AN-9, CI-994, Entinostat/MS-275, SNDX-275, mocetinostat/MGCD0103(N-(2-aminophenyl)-4-((4-pyridin-3-ylpyrimidin-2-ylamino)methyl)benzamide),m-carboxycinnamic acid, bishydroxamic acid, suberic bishydroxamic acid,oxamflatin, ABHA, SB-55629, pyroxamide, propenamides, aroyl pyrrolylhydroxamides, and LAQ824(((E)-N-hydroxy-3-[4-[[2-hydroxyethyl-[2-(1H-indol-3-yl)ethyl]amino]methyl]phenyl]prop-2-enamide).In other embodiments, the HDAC inhibitor is a largazole derivative. Incertain embodiments, the HDAC inhibitor is aN-hydroxypyrimidine-5-carboxamide, wherein the HDAC inhibitor comprisesan azabicyclo-hexane, wherein the HDAC inhibitor comprises afluoroquinoline group, or wherein the HDAC inhibitor is anon-piperidine-containing pyrimidine hydroxamic acid derivative.

In certain embodiments, the DNA virus is a herpes virus. In someembodiments, the herpes virus is an Epstein-Barr virus.

In some embodiments, the HDAC inhibitor and the antiviral agent areco-formulated. In certain embodiments, the co-formulation comprises aunit dose of no greater than 80 mg of the HDAC inhibitor and no greaterthan 1500 mg of the antiviral agent.

In certain embodiments, the HDAC inhibitor can penetrate the blood brainbarrier.

In some embodiments, the method further comprises administering anadditional agent. In certain embodiments, the additional agent is anantiviral agent, a HDAC inhibitor, or a chemotherapeutic.

In certain embodiments, the virally-induced condition is a cancerouscondition, an inflammatory condition, an autoimmune condition, anallergic condition, or a skin condition. In some embodiments, thevirally-induced condition is a lymphoma, chronic lymphocytic leukemia,nasopharyngeal carcinoma, gastric cancer, Kaposi's sarcoma, rheumatoidarthritis, systemic lupus erythematosus, or multiple sclerosis.

Also provided herein, in an additional aspect, is a method for treatingand/or preventing a viral condition, a virally-induced condition, or aninflammatory condition comprising administering a HDAC inhibitor and anantiviral agent wherein the MW of the HDAC inhibitor is greater than 275g/mol. In some embodiments, the HDAC inhibitor is aN-hydroxypyrimidine-5-carboxamide, wherein the HDAC inhibitor comprisesan azabicyclo-hexane, wherein the HDAC inhibitor comprises afluoroquinoline group, or wherein the HDAC inhibitor is anon-piperidine-containing pyrimidine hydroxamic acid derivative. Incertain embodiments, the HDAC inhibitor is JNJ-26481585(N-hydroxy-2-(4-((((1-methyl-1H-indol-3-yl)methyl)amino)methyl)piperidin-1-yl)pyrimidine-5-carboxamide),R306465/JNJ-16241199 (N-hydroxy-5-(4-(naphthalen-2-ylsulfonyl)piperazin-1-yl)pyrimidine-2-carboxamide), or CHR-3996(2-(6-{[(6-Fluoroquinolin-2-yl)methyl]amino}-3-azabicyclo[3.1.0]hex-3-yl)-N-hydroxypyrimidine-5-carboxamide).In other embodiments, the HDAC inhibitor is a largazole derivative.

In some embodiments, the viral or virally induced condition is caused bya herpes virus. In certain embodiments, the herpes virus is anEpstein-Barr virus.

In some embodiments, the HDAC inhibitor and the antiviral agent areadministered simultaneously. In certain embodiments, the HDAC inhibitorand the antiviral agent are administered orally.

In some embodiments, the HDAC inhibitor is administered at 0.01-1 mg/kgper dose. In certain embodiments, 1, 2, 3, or 4 doses are administereddaily. In some embodiments, the total daily dosage of the HDAC inhibitoris no greater than about 200 mg. In certain embodiments, the antiviralagent is valganciclovir and is administered at dose of 500-1500 mg/dose.

In some embodiments, the HDAC inhibitor and the antiviral agent areco-formulated. In certain embodiments, the co-formulation comprises aunit dose of no greater than 80 mg of the HDAC inhibitor and no greaterthan 1500 mg of the antiviral agent. In some embodiments, the antiviralagent is valganciclovir, and the valganciclovir is a timed release orslow release oral formulation.

In certain embodiments, the viral condition or virally-induced conditionis caused by a human immunodeficiency virus, a herpes virus, aparvovirus, a coxsackie virus, a Human T-lymphotropic virus, a BK virus,or a hepatitis virus. In some embodiments, the viral condition orvirally-induced condition is caused by a retrovirus or a herpes virus.In certain embodiments, the herpes virus is a herpes simplex virus, aherpes genitalis virus, a varicella zoster virus, an Epstein-Barr virus,a human herpes virus 6, a herpes virus type 1, herpes virus type 2, ahuman herpes virus type 8, or a cytomegalovirus. In some embodiments,the herpes virus is the Epstein-Barr virus. In certain embodiments, theviral condition or virally-induced condition is caused by a DNA virus.In some embodiments, the viral condition or virally-induced condition isnot caused by a retrovirus.

In certain embodiments, the virally-induced condition is a cancer, aninflammatory condition, an allergic condition, an autoimmune condition,or a skin condition. In some embodiments, the virally-induced conditionis lymphoma, chronic lymphocytic leukemia, nasopharyngeal carcinoma,gastric cancer, Kaposi's sarcoma, rheumatoid arthritis, systemic lupuserythematosus, or multiple sclerosis. In certain embodiments, thevirally-induced condition is not sepsis or viremia. In some embodiments,the inflammatory condition is an autoimmune condition, and allergiccondition, or a skin condition.

In certain embodiments, the HDAC inhibitor can penetrate the blood brainbarrier. In some embodiments, the antiviral agent is a HIV drug, aHerpes drug, a CMV drug, or a hepatitis drug. In certain embodiments,the antiviral agent is acyclovir, ganciclovir or valganciclovir. In someembodiments, the antiviral agent is not a heat shock protein inhibitor,an immunosuppressant, an antibiotic, a glucocorticoid, a non-steroidalanti-inflammatory drug, a Cox-2-specific inhibitor or a TNF-α bindingprotein. In certain embodiments, the antiviral agent is not a Hsp90inhibitor, tacrolimus, cyclosporin, rapamycin (sirolimus), methotrexate,cyclophosphamide, azathioprine, mercaptopurine, mycophenolate, FTY720,levofloxacin, amoxycillin, prednisone, cortisone acetate, prednisolone,methylprednisolone, dexamethasone, betamethasone, triamcinolone,beclometasone, fludrocortisone acetate, deoxycorticosterone acetate,aldosterone, salicylates, arylalkanoic acids, a 2-arylpropionic acid, aN-arylanthranilic acid, an oxicam, a coxib, a sulphonanilide,valdecoxib, celecoxib, rofecoxib, leflunomide, gold thioglucose, goldthiomalate, aurofin, sulfasalazine, hydroxychloroquinine, minocycline,infliximab, etanercept, adalimumab, abatacept, anakinra, interferon-β,interferon-γ, interleukin-2, an allergy vaccine, an antihistamine, anantileukotriene, a beta-agonist, theophylline, or an anticholinergic.

In some embodiments, the method further comprises administering anadditional agent. In certain embodiments, the additional agent is anantiviral agent, a HDAC inhibitor, or a chemotherapeutic.

Further provided herein, in one aspect, is a method for treating and/orpreventing a viral condition, a virally-induced condition, or aninflammatory condition comprising administering a HDAC inhibitor and anantiviral agent, wherein the HDAC inhibitor is a heterocyclic hydroxamicacid derivative. In some embodiments, the HDAC inhibitor is a pyrimidinehydroxamic acid derivative. In certain embodiments, the HDAC inhibitoris a N-hydroxypyrimidine-5-carboxamide, wherein the HDAC inhibitorcomprises an azabicyclo-hexane, wherein the HDAC inhibitor comprises afluoroquinoline group, or wherein the HDAC inhibitor does not comprise apiperidine group. In some embodiments, the HDAC inhibitor isJNJ-26481585(N-hydroxy-2-(4-((((1-methyl-1H-indol-3-yl)methyl)amino)methyl)piperidin-1-yl)pyrimidine-5-carboxamide),R306465/JNJ-16241199(N-hydroxy-5-(4-(naphthalen-2-ylsulfonyl)piperazin-1-yl)pyrimidine-2-carboxamide),or CHR-3996(2-(6-{[(6-Fluoroquinolin-2-yl)methyl]amino}-3-azabicyclo[3.1.0]hex-3-yl)-N-hydroxypyrimidine-5-carboxamide).

In some embodiments, the HDAC inhibitor and the antiviral agent areadministered simultaneously. In certain embodiments, the HDAC inhibitorand the antiviral agent are administered orally. In some embodiments,the antiviral agent is valganciclovir. In certain embodiments, the HDACinhibitor is administered at 0.2-2 mg/kg per dose. In some embodiments,1, 2, 3, or 4 doses are administered daily. In certain embodiments, thetotal daily dosage of the HDAC inhibitor is no greater than about 100mg. In some embodiments, valganciclovir is administered at dose of 900mg/dose or less.

In certain embodiments, the HDAC inhibitor and the antiviral agent areco-formulated. In some embodiments, the co-formulation comprises a unitdose of no greater than 80 mg of the HDAC inhibitor and no greater than1500 mg of the antiviral agent.

In certain embodiments, the viral condition or virally-induced conditionis caused by a human immunodeficiency virus, a herpes virus, aparvovirus, a coxsackie virus, a Human T-lymphotropic virus, a BK virus,or a hepatitis virus. In specific embodiments, the herpes virus is aherpes simplex virus, a herpes genitalis virus, a varicella zostervirus, an Epstein-Barr virus, a human herpes virus 6, a herpes virustype 1, herpes virus type 2, a human herpes virus type 8, or acytomegalovirus.

In some embodiments, the viral condition or virally-induced condition iscaused by a DNA virus. In certain embodiments, the viral condition orvirally-induced condition is caused by an Epstein-Barr virus. In someembodiments, the viral condition or virally-induced condition is notcaused by a retrovirus. In certain embodiments, the virally-inducedcondition is a cancer, an inflammatory condition, an allergic condition,an autoimmune condition, or a skin condition. In some embodiments, thevirally-induced condition is lymphoma, chronic lymphocytic leukemia,nasopharyngeal carcinoma, gastric cancer, Kaposi's sarcoma, rheumatoidarthritis, systemic lupus erythematosus, or multiple sclerosis. Incertain embodiments, the virally-induced condition is not sepsis orviremia. In specific embodiments, the inflammatory condition is anautoimmune condition, and allergic condition, or a skin condition.

In some embodiments, the HDAC inhibitor can penetrate the blood brainbarrier.

In certain embodiments, the antiviral agent is a HIV drug, a Herpesdrug, a CMV drug, or a hepatitis drug. In some embodiments, theantiviral agent is acyclovir, ganciclovir, or valganciclovir. In certainembodiments, the antiviral agent is not a heat shock protein inhibitor,an immunosuppressant, an antibiotic, a glucocorticoid, a non-steroidalanti-inflammatory drug, a Cox-2-specific inhibitor or a TNF-α bindingprotein. In other embodiments, the antiviral agent is not a Hsp90inhibitor, tacrolimus, cyclosporin, rapamycin (sirolimus), methotrexate,cyclophosphamide, azathioprine, mercaptopurine, mycophenolate, FTY720,levofloxacin, amoxycillin, prednisone, cortisone acetate, prednisolone,methylprednisolone, dexamethasone, betamethasone, triamcinolone,beclometasone, fludrocortisone acetate, deoxycorticosterone acetate,aldosterone, salicylates, arylalkanoic acids, a 2-arylpropionic acid, aN-arylanthranilic acid, an oxicam, a coxib, a sulphonanilide,valdecoxib, celecoxib, rofecoxib, leflunomide, gold thioglucose, goldthiomalate, aurofin, sulfasalazine, hydroxychloroquinine, minocycline,infliximab, etanercept, adalimumab, abatacept, anakinra, interferon-β,interferon-γ, interleukin-2, an allergy vaccine, an antihistamine, anantileukotriene, a beta-agonist, theophylline, or an anticholinergic.

In some embodiments, the method further comprises administering anadditional agent. In certain embodiments, the additional agent is anantiviral agent, a HDAC inhibitor, or a chemotherapeutic therapy.

Also provided herein, in another aspect, is a method for treating and/orpreventing a viral condition or a virally-induced condition comprisingadministering a HDAC inhibitor wherein the HDAC inhibitor is apyrimidine hydroxamic acid derivative comprising an azabicyclo-hexane,or wherein the HDAC inhibitor is a non-piperidine-containing pyrimidinehydroxamic acid derivative. In some embodiments, the HDAC inhibitor is apyrimidine hydroxamic acid derivative comprising an azabicyclo-hexane.In certain embodiments, the HDAC inhibitor is anon-piperidine-containing pyrimidine hydroxamic acid derivative. In someembodiments, the HDAC inhibitor comprises a fluoroquinoline group.

In some embodiments, the method further comprises administering anantiviral agent. In certain embodiments, the HDAC inhibitor andantiviral agent are administered simultaneously. In some embodiments,the HDAC inhibitor is administered orally. In certain embodiments, theHDAC inhibitor is administered at 0.01-1 mg/kg per dose. In someembodiments, 1, 2, 3, or 4 doses are administered daily. In certainembodiments, the total daily dosage of the HDAC inhibitor is no greaterthan about 200 mg. In some embodiments, the antiviral agent isadministered at dose of 500-1500 mg/dose.

In certain embodiments, the HDAC inhibitor and the antiviral agent areco-formulated. In some embodiments, the co-formulation comprises a unitdose of no greater than 80 mg of the HDAC inhibitor and no greater than1500 mg of the antiviral agent. In certain embodiments, the antiviralagent is valganciclovir.

In some embodiments, the viral condition or virally-induced condition iscaused by a human immunodeficiency virus, a herpes virus, a parvovirus,a coxsackie virus, a Human T-lymphotropic virus, a BK virus, or ahepatitis virus. In specific embodiments, the herpes virus is a herpessimplex virus, a herpes genitalis virus, a varicella zoster virus, anEpstein-Barr virus, a human herpes virus 6, a herpes virus type 1,herpes virus type 2, a human herpes virus type 8, or a cytomegalovirus.In some embodiments, the herpes virus is the Epstein-Barr virus.

In certain embodiments, the viral condition or virally-induced conditionis caused by a DNA virus. In some embodiments, the viral condition orvirally-induced condition is not caused by a retrovirus. In certainembodiments, the virally-induced condition is a cancer, an inflammatorycondition, an allergic condition, an autoimmune condition, or a skincondition. In some embodiments, the virally-induced condition is alymphoma, chronic lymphocytic leukemia, nasopharyngeal carcinoma,gastric cancer, Kaposi's sarcoma, rheumatoid arthritis, systemic lupuserythematosus, or multiple sclerosis. In certain embodiments, thevirally-induced condition is not sepsis or viremia.

In some embodiments, the HDAC inhibitor can penetrate the blood brainbarrier. In certain embodiments, the antiviral agent is a HIV drug, aHerpes drug, a CMV drug, or a hepatitis drug. In some embodiments, theantiviral agent is acyclovir, ganciclovir or valganciclovir. In certainembodiments, the antiviral agent is not a heat shock protein inhibitor,an immunosuppressant, an antibiotic, a glucocorticoid, a non-steroidalanti-inflammatory drug, a Cox-2-specific inhibitor or a TNF-α bindingprotein. In other embodiments, the antiviral agent is not a Hsp90inhibitor, tacrolimus, cyclosporin, rapamycin (sirolimus), methotrexate,cyclophosphamide, azathioprine, mercaptopurine, mycophenolate, FTY720,levofloxacin, amoxycillin, prednisone, cortisone acetate, prednisolone,methylprednisolone, dexamethasone, betamethasone, triamcinolone,beclometasone, fludrocortisone acetate, deoxycorticosterone acetate,aldosterone, salicylates, arylalkanoic acids, a 2-arylpropionic acid, aN-arylanthranilic acid, an oxicam, a coxib, a sulphonanilide,valdecoxib, celecoxib, rofecoxib, leflunomide, gold thioglucose, goldthiomalate, aurofin, sulfasalazine, hydroxychloroquinine, minocycline,infliximab, etanercept, adalimumab, abatacept, anakinra, interferon-β,interferon-γ, interleukin-2, an allergy vaccine, an antihistamine, anantileukotriene, a beta-agonist, theophylline, or an anticholinergic.

In some embodiments, the method further comprises administering anadditional agent. In certain embodiments, the additional agent is anantiviral agent, a HDAC inhibitor, or a chemotherapeutic therapy.

Further provided herein, in another aspect, is a composition comprisinga (i) HDAC inhibitor and (ii) an antiviral agent wherein the HDACinhibitor is a pyrimidine hydroxamic acid derivative. In someembodiments, the HDAC inhibitor is JNJ-26481585(N-hydroxy-2-(4-((((1-methyl-1H-indol-3-yl)methyl)amino)methyl)piperidin-1-yl)pyrimidine-5-carboxamide),R306465/JNJ-16241199 (N-hydroxy-5-(4-(naphthalen-2-ylsulfonyl)piperazin-1-yl)pyrimidine-2-carboxamide), or CHR-3996(2-(6-{[(6-Fluoroquinolin-2-yl)methyl]amino}-3-azabicyclo[3.1.0]hex-3-yl)-N-hydroxypyrimidine-5-carboxamide).In some embodiments, the HDAC inhibitor is not m-carboxycinnamic acid,bishydroxamic acid, suberic bishydroxamic acid, Trichostatin A(7-[4-(dimethylamino)phenyl]-N-hydroxy-4,6-dimethyl-7-oxohepta-2,4-dienamide),SAHA (suberoyl anilide hydroxamic acid)/Vorinostat, oxamflatin, ABHA,SB-55629, pyroxamide, propenamides, aroyl pyrrolyl hydroxamides,Belinostat/PXD101, Papobinostat, LAQ824(((E)-N-hydroxy-3-[4-[[2-hydroxyethyl-[2-(1H-indol-3-yl)ethyl]amino]methyl]phenyl]prop-2-enamide),LBH589, or TSA. In certain embodiments, the HDAC inhibitor can penetratethe blood brain barrier.

In some embodiments, the antiviral agent is a HIV drug, a Herpes drug, aCMV drug, or a hepatitis drug. In certain embodiments, the antiviralagent is acyclovir, ganciclovir, or valganciclovir. In some embodiments,the antiviral agent is not a heat shock protein inhibitor, animmunosuppressant, an antibiotic, a glucocorticoid, a non-steroidalanti-inflammatory drug, a Cox-2-specific inhibitor or a TNF-α bindingprotein. In other embodiments, the antiviral agent is not a Hsp90inhibitor, tacrolimus, cyclosporin, rapamycin (sirolimus), methotrexate,cyclophosphamide, azathioprine, mercaptopurine, mycophenolate, FTY720,levofloxacin, amoxycillin, prednisone, cortisone acetate, prednisolone,methylprednisolone, dexamethasone, betamethasone, triamcinolone,beclometasone, fludrocortisone acetate, deoxycorticosterone acetate,aldosterone, salicylates, arylalkanoic acids, a 2-arylpropionic acid, aN-arylanthranilic acid, an oxicam, a coxib, a sulphonanilide,valdecoxib, celecoxib, rofecoxib, leflunomide, gold thioglucose, goldthiomalate, aurofin, sulfasalazine, hydroxychloroquinine, minocycline,infliximab, etanercept, adalimumab, abatacept, anakinra, interferon-β,interferon-γ, interleukin-2, an allergy vaccine, an antihistamine, anantileukotriene, a beta-agonist, theophylline, or an anticholinergic.

In certain embodiments, the composition further comprises an additionalagent. In some embodiments, the additional agent is an antiviral agent,a HDAC inhibitor, or a chemotherapeutic drug.

In some embodiments, the HDAC inhibitor and the antiviral agent are inan oral formulation. In certain embodiments, the HDAC inhibitor ispresent at 0.01-1 mg/kg per dose. In some embodiments, the oralformulation comprises a unit dose of no greater than 80 mg of the HDACinhibitor and no greater than 1500 mg of the antiviral agent.

Additionally provided herein, in a further aspect, is a method fortreating and/or preventing a virus-induced inflammatory condition in asubject comprising administering a viral inducing agent and an antiviralagent to the subject, thereby treating and/or preventing theinflammatory condition. In some embodiments, the virus is a member ofthe herpes virus family, human immunodeficiency virus, parvovirus, orcoxsackie virus. In certain embodiments, the member of the herpes virusfamily is herpes simplex virus, herpes genitalis virus, varicella zostervirus, Epstein-Barr virus, human herpesvirus 6, or cytomegalovirus. Insome embodiments, the member of the herpes virus family is Epstein-Barrvirus, cytomegalovirus, or human herpesvirus 6.

In certain embodiments, the inflammatory condition is an autoimmunecondition. In some embodiments, the autoimmune condition is rheumatoidarthritis, multiple sclerosis, Sjogren's syndrome, systemic lupuserythematosus, autoimmune hepatitis, autoimmune thyroiditis,hemophagocytic syndrome, diabetes, Crohn's condition, ulcerativecolitis, psoriasis, psoriatic arthritis, idiopathic thrombocytonpenicpupura, polymyositis, dermatomyositis, myasthenia gravis, autoimmunethryroiditis, Evan's syndrome, autoimmune hemolytic anemia, aplasticanemia, autoimmune neutropenia, scleroderma, Reiter's syndrome,ankylosing spondylitis, pemphnigus, pemphigoid or autoimmune hepatitis.In certain embodiments, the inflammatory condition is an allergiccondition. In some embodiments, the inflammatory condition is a skincondition. In some embodiments, the inflammatory condition is associatedwith coronary artery condition or peripheral artery condition. Incertain embodiments, the inflammatory condition is retinitis,pancreatitis, cardiomyopathy, pericarditis, colitis, glomerulonephritis,lung inflammation, esophagitis, gastritis, duodenitis, ileitis,meningitis, encephalitis, encephalomyelitis, transverse myelitis,cystitis, urethritis, mucositis, lymphadenitis, dermatitis, hepatitis,osteomyelitis, or herpes zoster. In some embodiments, the inflammatorycondition is atherosclerosis. In certain embodiments, the virus iscytomegalovirus or herpes simplex virus.

In some embodiments, the viral inducing agent is one or more of achemotherapeutic drug, HDAC inhibitor, or DNA demethylating agent. Incertain embodiments, the HDAC inhibitor is butyrate or MS-275.

In some embodiments, the viral inducing agent can penetrate the bloodbrain barrier. In certain embodiments, the viral inducing agentcomprises arginine butyrate. In some embodiments, the antiviral agent isganciclovir or valganciclovir.

In certain embodiments, the method further comprises administering anadditional agent. In some embodiments, the additional agent comprises avaccine. In certain embodiments, the vaccine comprises myelin basicprotein and the condition is multiple sclerosis. In some embodiments,the vaccine comprises an antigen and the condition is diabetes. Incertain embodiments, the additional agent is aspirin, naproxen,ibuprofen, or a statin. In some embodiments, the inflammatory conditionis an autoimmune condition and the additional agent is cyclosporine,azathiorprine, methotrexate, cyclophosphamide, FK506, tacrolimus,monoclonal antibody, anti-T cell monoclonal antibody, anti-B cellmonoclonal antibody, IL-2 receptor antibody, or a TNF inhibitor. Incertain embodiments, the monoclonal antibody is an anti-B cell antibody.In some embodiments, the anti-B cell antibody is anti-CD20. In certainembodiments, the anti-T cell antibody is an anti-CD3 antibody. In someembodiments, the anti-CD3 antibody is OKT3. In certain embodiments, theTNF inhibitor is infliximab (Remicade™), etanercept (Enbrel™),Adalimumab (Humira™), or an anti-IL-6 antibody.

In certain embodiments, the inflammatory condition is atherosclerosisand the additional agent is a lipid lowering agent. In some embodiments,the lipid lowering agent is rosuvastatin, atorvastatin, simvastatin, orlovastatin. In certain embodiments, the inflammatory condition ismultiple sclerosis and the additional agent is mitoxantrone, cladribine,or Campath antibody. In some embodiments, the viral inducing agent isadministered to the subject before the antiviral agent.

INCORPORATION BY REFERENCE

All publications, patents, and patent applications mentioned in thisspecification are herein incorporated by reference to the same extent asif each individual publication, patent, or patent application wasspecifically and individually indicated to be incorporated by reference.

The following are also incorporated by reference: U.S. Pat. Nos.6,677,302; 7,399,787; US 2009/0270497; US 2010/0093824; US 2010/0152155;WO1998/04290; WO 2004/113336; WO 2008/097654; Moffat, D. et al,Discovery of2-(6-{[(6-Fluoroquinolin-2-yl)methyl]amino}bicyclo[3.1.0]hex-3-yl)-N-hydroxypyrimidine-5-carboxamide(CHR-3996), a Class I Selective Orally Active HDAC inhibitor, Journal ofMedicinal Chemistry (2010), 53, 8663-8678; Glaser, K B, HDAC inhibitors:Clinical update and mechanisms-based potential, Biochem. Pharmacol.(2007); Ghosh, S. K., et al. 2007 Blood Cells, Molecules, and Diseases38:57-65.

BRIEF DESCRIPTION OF THE DRAWINGS

The novel features of the invention are set forth with particularity inthe appended claims. A better understanding of the features andadvantages of the present invention will be obtained by reference to thefollowing detailed description that sets forth illustrative embodiments,in which the principles of the invention are utilized, and theaccompanying drawings of which:

FIGS. 1A-1B illustrate a CT scan of tumor reduction after treatment witharginine butyrate (AB) and GCV. FIG. 1A and FIG. 1B show pre- andpost-treatment, respectively.

FIGS. 2A-2D illustrate results from toxicity assays withanti-herpesvirus drugs ganciclovir (GCV) and penciclovir (PCV). FIG. 2Aand FIG. 2B show results from various concentrations of GCV and PCV.

FIG. 2C and FIG. 2D show results from GCV and PCV used in combinationwith NaB.

FIGS. 3A-3C illustrate results from analysis of efficacy of anti-viralsusing short chain fatty acids as inducing agents. FIG. 3A shows cellcount results after using NaB. FIG. 3B shows results from VA.

FIG. 3C shows fold of TK expression induced.

FIGS. 4A-4B illustrate results from analysis of efficacy of anti-viralsusing hydroxamic acids as inducing agents. FIG. 4A shows cell countresults after using scriptaid. FIG. 4B shows fold of TK expressioninduced.

FIGS. 5A-5B illustrate results from analysis of efficacy of anti-viralsusing SAHA (Vorinostat) as an inducing agent. FIG. 5A shows cell countresults after using SAHA. FIG. 5B shows fold of TK expression induced.

FIGS. 6A-6B illustrate results from analysis of efficacy of anti-viralsusing LHB589 (Panobinostat) as an inducing agent. FIG. 6A shows cellcount results after using LHB589. FIG. 6B shows fold of TK expressioninduced.

FIG. 7 illustrates results from analysis of efficacy of anti-viralsusing PXD101, which induced a high level of TK expression at the 5 μMconcentration.

FIG. 8 illustrates results from analysis of efficacy of anti-viralsusing oxamflatin as an inducing agent. FIG. 8 shows cell count resultsafter using oxamflatin.

FIG. 9 illustrates results from analysis of efficacy of anti-viralsusing a cyclic tetrapeptide as an inducing agent. FIG. 9 shows cellcount results after using apicidin.

FIGS. 10A-10C illustrate results from analysis of efficacy ofanti-virals using a benzamide (MS-275) as an inducing agent. FIG. 10Ashows cell count results after using MS-275. FIG. 10B shows fold of TKexpression induced. FIG. 10C shows cell count results after treatingcells in combination for shorter time periods.

FIGS. 11A-11B illustrate chemical structures of largazole compoundsused. Shown in both FIG. 11A and FIG. 11B.

FIGS. 12A-12F illustrate results from analysis of efficacy ofanti-virals using largazoles as an inducing agent. FIGS. 12A, 12B, 12C,12D and 12E show cell count results after using various largazolecompounds. FIG. 12F shows fold of TK expression induced from variouslargazole compounds.

FIG. 13 illustrates results from treatment of an HIV-1-infected monocytecell line with combination therapy. Viral release (p24 release) wasmeasured through optical density (OD) measurement.

FIG. 14 illustrates results from treatment of an HIV-1-infected monocytecell line with combination therapy. Viral release (p24 release) wasmeasured through optical density (OD) measurement and then convertedinto pg of protein.

FIG. 15 is a flow diagram of one embodiment of the provided invention.

DETAILED DESCRIPTION OF THE INVENTION

Provided herein are methods and compositions for treating and/orpreventing viral conditions, virally-induced conditions, or inflammatoryconditions in a subject. The condition can be associated with latentviral infections. The methods can comprise the steps of administering aviral inducing agent and an antiviral agent to the subject. The methodcan comprise steps of administering a viral inducing agent, an antiviralagent, and one or more additional agents to a subject. The methodsinclude the co-administration of an oral HDAC inhibitor and an antiviralagent, either in the same or separate formulations.

The methods and compositions provided can be used to treat and/orprevent infection by any of the viruses described herein. The methodsand compositions can be used to treat and/or prevent any of theinflammatory conditions described herein. Any of the viral inducingagents and/or antiviral agents described herein can be used in themethods and compositions of the provided invention. The viral inducingagent can be an HDAC inhibitor. The HDAC inhibitor can be a pyrimidinehydroxamic acid derivative.

One or more additional agents described herein can be administered to asubject. An additional agent can be selected for administration based onthe type of condition the subject has or is suspected of having.

Another aspect of the present invention relates to formulations, routesof administration and effective doses for pharmaceutical compositionscomprising an agent or combination of agents, e.g., viral inducingagents, antiviral agents, or one or more additional agents. A viralinducing agent, antiviral agent, or one or more additional agents can beadministered to a subject in separate pharmaceutical compositions or canbe co-formulated in a single pharmaceutical composition.

Also provided are methods relating to dosing schedules for administeringa viral inducing agent, antiviral agent, or one or more additionalagents. One or more pharmaceutical compositions can be administered to asubject by “pulsed administration” over a period of time.

Overview

Like all other herpes viruses, EBV has two stages of replication, thelytic and the latent. Soon after primary infection, immunologicalsurveillance by the host forces EBV to enter the latent state ofinfection, where only few selected genes are expressed. EBV maintainsthis latent state in all EBV-associated tumors. Conventional anti-herpesvirus drugs, such as ganciclovir, acyclovir, etc., fail to act on theselatently-infected cells because the viral enzyme thymidine kinase (TK)or protein kinase (PK), which is necessary for the conversion of theprodrugs to their toxic metabolites, is not expressed inlatently-infected cells. Provided herein, in some embodiments, is acombination treatment wherein lytic replication is induced and antiviraldrugs are administered concurrently.

Previous studies using patient-derived cells in vitro, and also fromphase I/II clinical studies on a series of patients with EBV-associatedlymphomas, have clearly shown the great promise of this combinationtherapy approach. Strong epidemiological association of Epstein-BarrVirus (EBV) with various human lymphoid malignancies and in vitrostudies demonstrating tumorigenic activity of many EBV latent geneproducts suggest a causal relationship between EBV and these diseases.However, as EBV maintains a latent state of infection in theselymphomas, typical anti-herpes viral drugs, such as the nucleosideanalogs ganciclovir (GCV) or acyclovir, are ineffective as thesepro-drugs require expression of a lytic phase EBV protein, thymidinekinase (TK) or protein kinase (EBV-PK), for their activity. Therefore,selective induction of EBV lytic-phase gene expression in lymphoma cellsthat harbor latent EBV, coupled with simultaneous exposure to antiviraldrugs, has been advanced as promising targeted therapy, because ofresulting targeting of cytotoxicity to the EBV-infected tumor cells.

A variety of agents including short-chain fatty acids andchemotherapeutic drugs, have been used to induce EBV lytic-phaseinfection in cultured cells, but these in vitro studies have generallynot resulted in clinical application. For instance, arginine butyrateand GCV has successfully been used to treat EBV-positive lymphoidmalignancies in a recent Phase I/II clinical trial. In this study of 15patients with relapsed or refractory EBV-positive lymphoid tumors, 4patients achieved complete tumor remissions and 6 patients partial tumorremissions. However, the rapid metabolism of butyrate requirescontinuous IV administration of high doses. Butyrate has pan-HDACinhibitory activity, and it has been established that this activity isresponsible for the induction of the EBV-TK protein. HDAC inhibitorshave been shown to induce both EBV-TK and EBV-PK in EBV infected tumors.

In recent years, several potent HDAC inhibitors (HDACi) have been testedin the clinic as anticancer agents. In certain instances, HDACinhibitors, including some new, highly-potent compounds, induce EBVlytic phase gene expression and kill EBV-infected cells in combinationwith antiviral drugs. In some instances, HDAC inhibitors induce lyticphase gene expression in viruses and kill virus-infected cells incombination with antiviral drugs. HDAC inhibitors include, but are notlimited to, short-chain fatty acids (sodium butyrate and valproic acid),hydroxamic acids (Oxamflatin, Scriptaid, CHR-3996, Suberoyl anilidehydroxamic acid (SAHA), Panobinostat (LBH589) and Belinostat (PXD101)),the benzamide MS275, cyclic tetrapeptide Apicidin, and newly-identifiedHDAC inhibitor Largazole, which was originally isolated from a marinecyanobacterium. In any of the embodiments herein, the HDAC inhibitor ispreferably suitable for oral administration.

Methods and Compositions

In one aspect, provided herein are methods for treating and/orpreventing a viral condition, a virally-induced condition, or aninflammatory condition. In some embodiments, the condition is associatedwith a latent viral infection. In certain embodiments, the methodscomprise administering a viral inducing agent (e.g., an HDAC inhibitor)and an antiviral agent. In some embodiments, the methods compriseadministering an HDAC inhibitor and an antiviral agent. In certainembodiments, the HDAC inhibitor and the antiviral agent areco-formulated. In some embodiments, the methods comprise furtheradministering an additional viral inducing agent. In other embodiments,the methods comprise further administering an additional antiviralagent. In some embodiments, the methods comprise administeringadditional individual doses of the viral inducing agent and/or theantiviral agent.

Further provided herein are methods for treating and/or preventing aviral condition, a virally-induced condition, or an inflammatorycondition comprising administering an HDAC inhibitor. In someembodiments, the HDAC inhibitor is a pyrimidine hydroxamic acidderivative. In certain embodiments, the HDAC inhibitor is a pyrimidinehydroxamic acid derivative not containing a piperidine, or a pyrimidinehydroxamic acid derivative comprising an azabicyclohexane. In certainembodiments, the methods further comprise administering an antiviralagent. In some embodiments, the HDAC inhibitor and the antiviral agentare co-formulated.

Also provided herein are methods for treating and/or preventing a viralor virally-induced condition, or an inflammatory condition comprisingadministering an HDAC inhibitor and an antiviral agent. In someembodiments, the viral or virally-induced condition is caused by a DNAvirus. In certain embodiments, the HDAC inhibitor is administered at adose of less than 2 mg/kg per dose. In some embodiments, the HDACinhibitor is administered at a dose of less than 20 mg/kg, less than 19mg/kg, less than 18 mg/kg, less than 17 mg/kg, less than 16 mg/kg, lessthan 15 mg/kg, less than 14 mg/kg, less than 13 mg/kg, less than 12mg/kg, less than 11 mg/kg, less than 10 mg/kg, less than 9 mg/kg, lessthan 8 mg/kg, less than 7 mg/kg, less than 6 mg/kg, less than 5 mg/kg,less than 4 mg/kg, less than 3 mg/kg, less than 2 mg/kg, less than 1mg/kg, less than 0.5 mg/kg, less than 0.2 mg/kg, or less than 0.1 mg/kg.

Further provided herein are methods for treating and/or preventing aviral condition, a virally-induced condition, or an inflammatorycondition comprising administering an HDAC inhibitor and an antiviralagent wherein the HDAC inhibitor has a molecular weight of greater than275 g/mol. In some embodiments, the HDAC inhibitor has a molecularweight of greater than 200 g/mol, greater than 225 g/mol, greater than250 g/mol, greater than 275 g/mol, greater than 300 g/mol, greater than325 g/mol, greater than 350 g/mol, greater than 375 g/mol, greater than400 g/mol, greater than 425 g/mol, greater than 450 g/mol, greater than475 g/mol, greater than 500 g/mol, greater than 525 g/mol, greater than550 g/mol, greater than 575 g/mol, greater than 600 g/mol, greater than625 g/mol, greater than 650 g/mol, greater than 675 g/mol, greater than700 g/mol, greater than 725 g/mol, greater than 750 g/mol, greater than775 g/mol, greater than 800 g/mol, greater than 850 g/mol, greater than900 g/mol, greater than 950 g/mol, or greater than 1000 g/mol. Incertain embodiments, the HDAC inhibitor has a molecular weight of lessthan 200 g/mol, less than 225 g/mol, less than 250 g/mol, less than 275g/mol, less than 300 g/mol, less than 325 g/mol, less than 350 g/mol,less than 375 g/mol, less than 400 g/mol, less than 425 g/mol, less than450 g/mol, less than 475 g/mol, less than 500 g/mol, less than 525g/mol, less than 550 g/mol, less than 575 g/mol, less than 600 g/mol,less than 625 g/mol, less than 650 g/mol, less than 675 g/mol, less than700 g/mol, less than 725 g/mol, less than 750 g/mol, less than 775g/mol, less than 800 g/mol, less than 850 g/mol, less than 900 g/mol,less than 950 g/mol, or less than 1000 g/mol. In some embodiments, theHDAC inhibitor has a molecular weight of less than 500 g/mol and morethan 250 g/mol. In other embodiments, the HDAC inhibitor has a molecularweight of less than 400 g/mol and more than 300 g/mol. In someinstances, the HDAC inhibitor is not butyric acid or arginine butyrate.

Also provided herein are methods for treating and/or preventingEpstein-Barr virus (EBV) associated lymphoma. In some embodiments, themethods comprise administering an HDAC inhibitor and acyclovir and/organciclovir and/or valganciclovir.

Further provided here are methods for treating and/or preventing avirus-induced inflammatory condition. In some embodiments, the methodscomprise administering a viral inducing agent and an antiviral agent. Incertain embodiments the virus is a member of the herpes family (e.g.,herpes simplex virus, herpes genitalis virus, varicella zoster virus,Epstein-Barr virus, human herpes virus 6, or cytomegalovirus), humanimmunodeficiency virus, parvovirus, or coxsackie virus.

In another aspect, provided herein are compositions comprising an HDACinhibitor and an antiviral agent. In certain embodiments, the HDACinhibitor is a pyrimidine hydroxamic acid derivative. In someembodiments, the HDAC inhibitor is JNJ-26481585, JNJ-16241199, orCHR-3996. In some embodiments, the antiviral agent is acyclovir,ganciclovir, or valganciclovir. In certain embodiments, the compositioncomprises an additional agent. In some embodiments, the additional agentis a antiviral agent, an HDAC inhibitor, or a chemotherapeutic agent. Incertain embodiments, the compositions are formulated as a capsule, gel,tablet, solution, or suspension. In some embodiments, the compositionsare formulated for oral administration. In other embodiments, thecompositions are formulated for parenteral administration. In someembodiments, the compositions are formulated for intravenous,intraperitoneal, oral, subcutaneous, intrathecal, or intratumoraladministration. In certain embodiments, the compositions are formulatedfor administration at the site of a viral infection. In someembodiments, the compositions are formulated for modified release of theHDAC inhibitor and the antiviral agent. In specific embodiments, theHDAC inhibitor is dissolved before the antiviral agent is dissolved. Inother specific embodiments, the HDAC inhibitor is dissolved after theantiviral agent is dissolved. In some embodiments, the compositions areformulated for once daily administration. In other embodiments, thecompositions are formulated for twice daily, thrice daily, four timesdaily, once every other day, once weekly, once bi-weekly, or monthly.

Definitions

The terms “viral,” “virus-associated,” and “virally-induced” withreference to disorders are used interchangeably throughout the instantspecification.

The term “obtaining” as in “obtaining the composition” is intended toinclude purchasing, synthesizing, or otherwise acquiring the composition(or agent(s) of the composition).

The terms “comprises”, “comprising”, are intended to have the broadmeaning ascribed to them and can mean “includes”, “including” and thelike.

The term “subject”, “patient” or “individual” are used interchangeablyherein and refer to mammals and non-mammals, e.g., suffering from adisorder described herein. Examples of mammals include, but are notlimited to, any member of the Mammalian class: humans, non-humanprimates such as chimpanzees, and other apes and monkey species; farmanimals such as cattle, horses, sheep, goats, swine; domestic animalssuch as rabbits, dogs, and cats; laboratory animals including rodents,such as rats, mice and guinea pigs, and the like. Examples ofnon-mammals include, but are not limited to, birds, fish and the like.In one embodiment of the methods and compositions provided herein, themammal is a human.

The terms “treat,” “treating” or “treatment,” and other grammaticalequivalents as used herein, include alleviating, inhibiting or reducingsymptoms, reducing or inhibiting severity of, reducing incidence of,prophylactic treatment of, reducing or inhibiting recurrence of,delaying onset of, delaying recurrence of, abating or ameliorating adisease or condition symptoms, ameliorating the underlying metaboliccauses of symptoms, inhibiting the disease or condition, e.g., arrestingthe development of the disease or condition, relieving the disease orcondition, causing regression of the disease or condition, relieving acondition caused by the disease or condition, or stopping the symptomsof the disease or condition. The terms further include achieving atherapeutic benefit. By therapeutic benefit is meant eradication oramelioration of the underlying disorder being treated, and/or theeradication or amelioration of one or more of the physiological symptomsassociated with the underlying disorder such that an improvement isobserved in the patient.

The terms “prevent,” “preventing” or “prevention,” and other grammaticalequivalents as used herein, include preventing additional symptoms,preventing the underlying metabolic causes of symptoms, inhibiting thedisease or condition, e.g., arresting the development of the disease orcondition and are intended to include prophylaxis. The terms furtherinclude achieving a prophylactic benefit. For prophylactic benefit, thecompositions are optionally administered to a patient at risk ofdeveloping a particular disease, to a patient reporting one or more ofthe physiological symptoms of a disease, or to a patient at risk ofreoccurrence of the disease.

The terms “effective amount” or “therapeutically effective amount” asused herein, refer to a sufficient amount of at least one agent beingadministered which achieve a desired result, e.g., to relieve to someextent one or more symptoms of a disease or condition being treated. Incertain instances, the result is a reduction and/or alleviation of thesigns, symptoms, or causes of a disease, or any other desired alterationof a biological system. In certain instances, an “effective amount” fortherapeutic uses is the amount of the composition comprising an agent asset forth herein required to provide a clinically significant decreasein a disease. An appropriate “effective” amount in any individual caseis determined using any suitable technique, such as a dose escalationstudy.

The terms “administer,” “administering”, “administration,” and the like,as used herein, refer to the methods that are used to enable delivery ofagents or compositions to the desired site of biological action. Thesemethods include, but are not limited to oral routes, intraduodenalroutes, parenteral injection (including intravenous, subcutaneous,intraperitoneal, intramuscular, intravascular or infusion), topical andrectal administration. Administration techniques that in some instancesare employed with the agents and methods described herein include, e.g.,as discussed in Goodman and Gilman, The Pharmacological Basis ofTherapeutics (current edition), Pergamon; and Remington's,Pharmaceutical Sciences (current edition), Mack Publishing Co., Easton,Pa. In certain embodiments, the agents and compositions described hereinare administered orally. In some embodiments, the compositions describedherein are administered parenterally.

The term “pharmaceutically acceptable” as used herein, refers to amaterial that does not abrogate the biological activity or properties ofthe agents described herein, and is relatively nontoxic (i.e., thetoxicity of the material significantly outweighs the benefit of thematerial). In some instances, a pharmaceutically acceptable material isadministered to an individual without causing significant undesirablebiological effects or significantly interacting in a deleterious mannerwith any of the components of the composition in which it is contained.

The term “pharmaceutically acceptable excipient,” as used herein, refersto carriers and vehicles that are compatible with the active ingredient(for example, a compound of the invention) of a pharmaceuticalcomposition of the invention (and preferably capable of stabilizing it)and not deleterious to the subject to be treated. For example,solubilizing agents that form specific, more soluble complexes with thecompounds of the invention can be utilized as pharmaceutical excipientsfor delivery of the compounds. Suitable carriers and vehicles are knownto those of extraordinary skill in the art. The term “excipient” as usedherein will encompass all such carriers, adjuvants, diluents, solvents,or other inactive additives. Suitable pharmaceutically acceptableexcipients include, but are not limited to, water, salt solutions,alcohol, vegetable oils, polyethylene glycols, gelatin, lactose,amylose, magnesium stearate, talc, silicic acid, viscous paraffin,perfume oil, fatty acid monoglycerides and diglycerides, petroethralfatty acid esters, hydroxymethyl-cellulose, polyvinylpyrrolidone, etc.The pharmaceutical compositions of the invention can also be sterilizedand, if desired, mixed with auxiliary agents, e.g., lubricants,preservatives, stabilizers, wetting agents, emulsifiers, salts forinfluencing osmotic pressure, buffers, colorings, flavorings and/oraromatic substances and the like, which do not deleteriously react withthe active compounds of the invention.

The invention can be understood more fully by reference to the followingdetailed description and illustrative examples, which are intended toexemplify non-limiting embodiments of the invention.

Viral Inducing Agents

The methods of the provided invention comprise use of one or morepharmaceutical compositions provided herein comprising an inducing agentto induce expression of a gene product in a virus-infected cell. Thegene product expressed can be a viral enzyme or a cellular enzyme oractivity that is largely expressed in virus-infected cells. Expressionproducts that can be targeted include enzymes involved with DNAreplication, for example, for repair or replication of the genome,assembly of complete virus particles, generation of viral membrane orwalls, RNA transcription or protein translation or combinations of theseactivities. Interference with these processes can be performed byinducing and then acting on an enzyme and, preferably, a critical enzymein the process. Inducing agents that can be used in the methods andcompositions of the provided invention are described, for example, inU.S. Pat. Nos. 6,197,743 and 6,677,302, which are herein incorporated byreference in their entireties.

Inducing agents according to the methods or compositions provided hereininclude, without limitation, short-chain fatty acid (SCFA) derivatives,histone deacetylase (HDAC) inhibitors, phorbol esters, anticanceragents, and cytokines. In some embodiments, the viral inducing agent isa chemotherapeutic drug, an HDAC inhibitor, or a DNA demethylatingagent.

In some embodiments, the inducing agent is a SCFA derivative. Examplesof SCFA inducing agents include propionic acid, butyric acid, succinicacid, fumaric acid monoethyl ester, dimethyl butyric acid,trifluorobutanol, chloropropionic acid, isopropionic acid,2-oxypentanoic acid, 2,2- or 3,3-dimethyl butyric acid, 2,2- or3,3-diethyl butyric acid, butyric acid ethyl ester, 2-methyl butanoicacid, fumaric acid, and amides and salts thereof. Other examples includemethoxy acetic acid, methoxy propionic acid, N-acetylglycine,mercaptoacetic acid, 1- or 2-methyl cyclopropane carboxylic acid,squaric acid, 2- or 3-phenoxy propionic acid, methoxy butyric acid,phenoxy acetic acid, 2- or 3-phenoxy butyric acid, phenyl acetic acid,phenyl propionic acid, 3-phenyl butyric acid, ethyl-phenyl acetic acid,4-chloro-2-phenoxy-2-propionic acid, n-dimethyl butyric acid glycineamide, o-benzoyl lactic acid, o-dimethyl butyric acid lactate, cinnamicacid, dihydrocinnamic acid (C₆ H₅ CHCH₃ COOH),alpha-methyl-dihydrocinnamic acid, thiophenoxy acetic acid, and amines,amides and salts of these chemicals. Useful amines and amides caninclude isobutylhydroxylamine, fumaric acid monoamide, fumaramide,succinamide, or isobutyramide.

In other embodiments, inducing agents include retinoic acid, retinol,cytosine arabinoside, phorbols such as the phorbol diester12-0-tetradecanoylphorbol 13-acetate (TPA), teleocidine B, indolealkaloids, cytotoxin, plant lectins from Streptomyces, glucocorticoidssuch as estrogen and progesterone, phytohemagglutinin (PHA), bryostatin,growth factors (e.g. PDGF, VEGF, EGF, FGF, NGF, TGF, BCGF), anti-sensenucleic acids (e.g. DNA, RNA or PNA), aptamers (nucleic acidoligonucleotides with secondary or tertiary structures which bind withhigh affinity and selectivity to a target molecule), erythropoietin(EPO), the interleukins (IL-1, IL-2, IL-3, etc.), cAMP and cAMP analogssuch as dibutyryl cAMP, activin, inhibin, steel factor, interferon, thebone morphogenic proteins (BMBs), hydroxyurea and dimethyl sulfoxide(DMSO). Other inducing agents include interferons (e.g. α-, β-,γ-interferon), cytokines such as tumor necrosis factor (TNF), cellreceptors, and growth factor antagonists, which may be purified orrecombinantly produced.

In some embodiments, the inducing agent is a anticancer agent. Incertain embodiments, the anticancer agent is a chemotherapeuticanticancer agent. Examples of chemotherapeutic anticancer agents includeNitrogen Mustards like bendamustine, chlorambucil, chlormethine,cyclophosphamide, ifosfamide, melphalan, prednimustine, trofosfamide;Alkyl Sulfonates like busulfan, mannosulfan, treosulfan; Ethylene Imineslike carboquone, thiotepa, triaziquone; Nitrosoureas like carmustine,fotemustine, lomustine, nimustine, ranimustine, semustine, streptozocin;Epoxides like etoglucid; Other Alkylating Agents like dacarbazine,mitobronitol, pipobroman, temozolomide; Folic Acid Analogues likemethotrexate, permetrexed, pralatrexate, raltitrexed; Purine Analogslike cladribine, clofarabine, fludarabine, mercaptopurine, nelarabine,tioguanine; Pyrimidine Analogs like azacitidine, capecitabine, carmofur,cytarabine, decitabine, fluorouracil, gemcitabine, tegafur; VincaAlkaloids like vinblastine, vincristine, vindesine, vinflunine,vinorelbine; Podophyllotoxin Derivatives like etoposide, teniposide;Colchicine derivatives like demecolcine; Taxanes like docetaxel,paclitaxel, paclitaxel poliglumex; Other Plant Alkaloids and NaturalProducts like trabectedin; Actinomycines like dactinomycin;Antracyclines like aclarubicin, daunorubicin, doxorubicin, epirubicin,idarubicin, mitoxantrone, pirarubicin, valrubicin, zorubincin; OtherCytotoxic Antibiotics like bleomycin, ixabepilone, mitomycin,plicamycin; Platinum Compounds like carboplatin, cisplatin, oxaliplatin,satraplatin; Methylhydrazines like procarbazine; Sensitizers likeaminolevulinic acid, efaproxiral, methyl aminolevulinate, porfimersodium, temoporfin; Protein Kinase Inhibitors like dasatinib, erlotinib,everolimus, zotarolimus, gefitinib, imatinib, lapatinib, nilotinib,pazonanib, sorafenib, sunitinib, temsirolimus; Other AntineoplasticAgents like alitretinoin, altretamine, amzacrine, anagrelide, arsenictrioxide, asparaginase, bexarotene, bortezomib, celecoxib, denileukindiftitox, estramustine, hydroxycarbamide, irinotecan, lonidamine,masoprocol, miltefosein, mitoguazone, mitotane, oblimersen,pegaspargase, pentostatin, romidepsin, sitimagene ceradenovec,tiazofurine, topotecan, tretinoin, vorinostat; Estrogens likediethylstilbenol, ethinylestradiol, fosfestrol, polyestradiol phosphate;Progestogens like gestonorone, medroxyprogesterone, megestrol;Gonadotropin Releasing Hormone Analogs like buserelin, goserelin,leuprorelin, triptorelin; Anti-Estrogens like fulvestrant, tamoxifen,toremifene; Anti-Androgens like bicalutamide, flutamide, nilutamide;Enzyme Inhibitors like aminoglutethimide, anastrozole, exemestane,formestane, letrozole, vorozole; Other Hormone Antagonists likeabarelix, degarelix; Immunostimulants like histamine dihydrochloride,mifamurtide, pidotimod, plerixafor, roquinimex, thymopentin;Immunosuppressants like everolimus, zotarolimus, gusperimus,leflunomide, mycophenolic acid, sirolimus; Calcineurin Inhibitors likeciclosporin, tacrolimus; Other Immunosuppressants like azathioprine,lenalidomide, methotrexate, thalidomide; and Radiopharmaceuticals likeiobenguane.

In certain embodiments, the viral inducing agent is a demethylatingagent. For example, demethylating agents include decitabine andazacytidine. In other embodiments, the inducing agent is a chemotherapydrug, such as cyclophosphamide, cisplatin, melphalan, doxorubicin,daunorubicin, vincristine, methotrexate, cytarabine, ifosfamide,etoposide, or rituximab.

In further embodiments, inducing agents include histone deacetylase(HDAC) inhibitors (including those of the hydroxamic acid class and thebenzamide class), DNA methyltransferase inhibitors, and proteasomeinhibitors. HDAC inhibitors, a class of compounds that interfere withthe function of histone deacetylase, include, without limitation,short-chain fatty acids (butyrate, phenylbutyrate, valproate, AN-9,etc., as described above), hydroxamic acids (for examplem-carboxycinnamic acid, bishydroxamic acid, suberic bishydroxamic acid,Trichostatin A(7-[4-(dimethylamino)phenyl]-N-hydroxy-4,6-dimethyl-7-oxohepta-2,4-dienamide),SAHA (suberoyl anilide hydroxamic acid)/Vorinostat, oxamflatin, ABHA,SB-55629, pyroxamide, propenamides, aroyl pyrrolyl hydroxamides,Belinostat/PXD101, Papobinostat, LAQ824(((E)-N-hydroxy-3-[4-[[2-hydroxyethyl-[2-(1H-indol-3-yl)ethyl]amino]methyl]phenyl]prop-2-enamide),LBH589, CHR-3996(2-(6-{[(6-Fluoroquinolin-2-yl)methyl]amino}-3-azabicyclo[3.1.0]hex-3-yl)-N-hydroxypyrimidine-5-carboxamide,TSA), Pivanex, spiruchostatins, cyclic tetrapeptides (for example,trapoxin A(cyclo((S)-phenylalanyl-(S)-phenylalanyl-(R)-pipecolinyl-(2S,9S)-2-amino-8-oxo-9,10-epoxydecanoyl),),trapoxin B(cyclo((S)-phenylalanyl-phenylalanyl-(R)-prolyl-2-amino-8-oxo-9,10-epoxydecanoyl)),HC-toxin, chlamydocin, diheteropeptin, WF-3161, Cyl-1, Cyl-2, azumamideA), cyclic peptides (for example, FK-228, FR901228), depsipeptides (forexample, romidepsin, FK228((E)-(1S,4S,10S,21R)-7[(Z)-ethylideno]-4,21-diisopropyl-2-oxa-12,13-dithia-5,8,20,23-tetraazabicyclo[8,7,6]-tricos-16-ene-3,6,9,22-pentanone),FK228 analogs and derivatives, largazole, largazole analogs andderivatives), peptide antibiotics (apicidin), benzamides (MS275(3-pyridinylmethyl [[4-[[(2-aminophenyl) amino]carbonyl]phenyl]methyl]carbamate,N-(2-Aminophenyl)-4-[N-(pyridine-3ylmethoxycarbonyl)aminomethyl]benzamide),CI994 (4-(Acetylamino)-N-(2-aminophenyl)benzamide), MGCD0103),electrophilic ketones (TPX, AOR, Depudecin), FR901375, nicotinamide, NADderivatives, Sirtinol, splitomycin, dihydrocoumarin, naphthopyranone,2-hydroxynaphthaldehydes, PCYC-0402, PCYC-0403, PCI-24781(3-(dimethylaminomethyl)-N-[2-[4-(hydroxycarbamoyl)phenoxy]ethyl]-1-benzofuran-2-carboxamide),depudecin, tubacin, organosulfur compounds, and dimethyl sulfoxide(DMSO). Other compounds which may also be administered as inducingagents, which include CHAPs, Scriptaid, Tubacin, JNJ16241199, A-161906,6-(3-Chlorophenylureido)caproic hydroxamic acid, SB939, ITF2357({6-[(diethylamino)methyl]-2-naphthyl}methyl{4-[(hydroxyamino)carbonyl]phenyl}carbamate), 4SC-201, AR-42, OPB-801,RG2833, CUDC-101, JNJ-26481585, MK0683 (suberoylanilide hydroxamicacid), M344(4-(Diethylamino)-N-[7-(hydroxyamino)-7-oxoheptyl]benzamide), BML-210(N-(2-aminophenyl)-N′-phenyl-octanediamide), dacinostat (NVP-LAQ824),PDX-101, BAY86-5274, SB939, droxinostat, and pivaloyloxymethyl butyrate.

In some embodiments, the viral inducing agent is an HDAC inhibitor. Insome embodiments, the HDAC inhibitor is a hydroxamic acid, for example,Vorinostat/suberoyl anilide hydroxamic acid (SAHA), JNJ-26481585(N-hydroxy-2-(4-((((1-methyl-1H-indol-3-yl)methyl)amino)methyl)piperidin-1-yl)pyrimidine-5-carboxamide),R306465/JNJ-16241199(N-hydroxy-5-(4-(naphthalen-2-ylsulfonyl)piperazin-1-yl)pyrimidine-2-carboxamide), CHR-3996(2-(6-{[(6-Fluoroquinolin-2-yl)methyl]amino}-3-azabicyclo[3.1.0]hex-3-yl)-N-hydroxypyrimidine-5-carboxamide),Belinostat/PXD101, Panobinostat/LBH-589, trichostatin A/TSA(7-[4-(dimethylamino)phenyl]-N-hydroxy-4,6-dimethyl-7-oxohepta-2,4-dienamide),ITF2357, CBHA, and Givinostat/ITF2357. In certain embodiments, the HDACinhibitor is a pyrimidine hydroxamic acid, for example, JNJ-26481585,JNJ-16241199, or CHR-3996. In other embodiments, the HDAC inhibitor is abenzamide, for example, CI-994, Entinostat/MS-275, SNDX-275, andmocetinostat/MGCD0103(N-(2-aminophenyl)-4-((4-pyridin-3-ylpyrimidin-2-ylamino)methyl)benzamide).

In some embodiments, the HDAC inhibitor is a hydroxamic acid derivative.In certain embodiments, the HDAC inhibitor is a pyrimidine hydroxamicacid. In some embodiments, the HDAC inhibitor is anon-piperidine-containing pyrimidine hydroxamic acid derivative. Incertain embodiments, the HDAC inhibitor comprises an azabicyclo-hexane.In other embodiments, the HDAC inhibitor comprises fluorine. In certainembodiments, the HDAC inhibitor comprises a fluoroquinoline group.

In one embodiment the HDAC inhibitor is JNJ-26481585(N-hydroxy-2-(4-((((1-methyl-1H-indol-3-yl)methyl)amino)methyl)piperidin-1-yl)pyrimidine-5-carboxamide).In a further embodiment, the HDAC inhibitor is R306465/JNJ-16241199(N-hydroxy-5-(4-(naphthalen-2-ylsulfonyl)piperazin-1-yl)pyrimidine-2-carboxamide).In a specific embodiment, the HDAC inhibitor is CHR-3996[2-(6-{[(6-Fluoroquinolin-2-yl)methyl]amino}-3-azabicyclo[3.1.0]hex-3-yl)-N-hydroxypyrimidine-5-carboxamide,or N-hydroxy2-(6-{[(6-Fluoroquinolin-2-yl)methyl]amino}-3-azabicyclo[3.1.0]hex-3-yl)-pyrimidine-5-carboxamide].

In another embodiment, the molecular weight of the HDAC inhibitor isgreater than 275 g/mol. In some embodiments, the HDAC inhibitor has amolecular weight of greater than 200 g/mol, greater than 225 g/mol,greater than 250 g/mol, greater than 275 g/mol, greater than 300 g/mol,greater than 325 g/mol, greater than 350 g/mol, greater than 375 g/mol,greater than 400 g/mol, greater than 425 g/mol, greater than 450 g/mol,greater than 475 g/mol, greater than 500 g/mol, greater than 525 g/mol,greater than 550 g/mol, greater than 575 g/mol, greater than 600 g/mol,greater than 625 g/mol, greater than 650 g/mol, greater than 675 g/mol,greater than 700 g/mol, greater than 725 g/mol, greater than 750 g/mol,greater than 775 g/mol, greater than 800 g/mol, greater than 850 g/mol,greater than 900 g/mol, greater than 950 g/mol, or greater than 1000g/mol. In certain embodiments, the HDAC inhibitor has a molecular weightof less than 200 g/mol, less than 225 g/mol, less than 250 g/mol, lessthan 275 g/mol, less than 300 g/mol, less than 325 g/mol, less than 350g/mol, less than 375 g/mol, less than 400 g/mol, less than 425 g/mol,less than 450 g/mol, less than 475 g/mol, less than 500 g/mol, less than525 g/mol, less than 550 g/mol, less than 575 g/mol, less than 600g/mol, less than 625 g/mol, less than 650 g/mol, less than 675 g/mol,less than 700 g/mol, less than 725 g/mol, less than 750 g/mol, less than775 g/mol, less than 800 g/mol, less than 850 g/mol, less than 900g/mol, less than 950 g/mol, or less than 1000 g/mol. In someembodiments, the HDAC inhibitor has a molecular weight of less than 500g/mol and more than 250 g/mol. In other embodiments, the HDAC inhibitorhas a molecular weight of less than 400 g/mol and more than 300 g/mol.

In a particular embodiment, the HDAC inhibitor is not m-carboxycinnamicacid, bishydroxamic acid, suberic bishydroxamic acid, Trichostatin A(7-[4-(dimethylamino)phenyl]-N-hydroxy-4,6-dimethyl-7-oxohepta-2,4-dienamide),SAHA (suberoyl anilide hydroxamic acid)/Vorinostat, oxamflatin, ABHA,SB-55629, pyroxamide, propenamides, aroyl pyrrolyl hydroxamides,Belinostat/PXD101, Papobinostat, LAQ824(((E)-N-hydroxy-3-[4-[[2-hydroxyethyl-[2-(1H-indol-3-yl)ethyl]amino]methyl]phenyl]prop-2-enamide),or LBH589.

In some embodiments, a viral inducing agent, for example an HDACinhibitor, penetrates the blood brain barrier. In certain embodiments, aviral inducing agent that penetrates the blood brain barrier comprisesarginine butyrate, SAHA, or CHR-3996.

In certain embodiments, the HDAC inhibitor is administered at a dose ofless than 400 mg/day. In some embodiments, the HDAC inhibitor isadministered at a dose of about 1 mg/day, about 2 mg/day, about 5mg/day, about 10 mg/day, about 15 mg/day, about 20 mg/day, about 25mg/day, about 30 mg/day, about 35 mg/day, about 40 mg/day, about 45mg/day, about 50 mg/day, about 60 mg/day, about 70 mg/day, about 80mg/day, about 90 mg/day, about 100 mg/day, about 120 mg/day, about 125mg/day, about 140 mg/day, about 150 mg/day, about 160 mg/day, about 175mg/day, about 180 mg/day, about 190 mg/day, about 200 mg/day, about 225mg/day, about 250 mg/day, about 275 mg/day, about 300 mg/day, about 325mg/day, about 350 mg/day, about 375 mg/day, about 400 mg/day, about 425mg/day, about 450 mg/day, about 475 mg/day, or about 500 mg/day. Incertain embodiments, the HDAC inhibitor is administered at a dose ofless than 1 mg/day, less than 2 mg/day, less than 5 mg/day, less than 10mg/day, less than 15 mg/day, less than 20 mg/day, less than 25 mg/day,less than 30 mg/day, less than 35 mg/day, less than 40 mg/day, less than45 mg/day, less than 50 mg/day, less than 60 mg/day, less than 70mg/day, less than 80 mg/day, less than 90 mg/day, less than 100 mg/day,less than 120 mg/day, less than 125 mg/day, less than 140 mg/day, lessthan 150 mg/day, less than 160 mg/day, less than 175 mg/day, less than180 mg/day, less than 190 mg/day, less than 200 mg/day, less than 225mg/day, less than 250 mg/day, less than 275 mg/day, less than 300mg/day, less than 325 mg/day, less than 350 mg/day, less than 375mg/day, less than 400 mg/day, less than 425 mg/day, less than 450mg/day, less than 475 mg/day, or less than 500 mg/day. In someembodiments, the HDAC inhibitor is administered at a dose of more than 1mg/day, more than 2 mg/day, more than 5 mg/day, more than 10 mg/day,more than 15 mg/day, more than 20 mg/day, more than 25 mg/day, more than30 mg/day, more than 35 mg/day, more than 40 mg/day, more than 45mg/day, more than 50 mg/day, more than 60 mg/day, more than 70 mg/day,more than 80 mg/day, more than 90 mg/day, more than 100 mg/day, morethan 120 mg/day, more than 125 mg/day, more than 140 mg/day, more than150 mg/day, more than 160 mg/day, more than 175 mg/day, more than 180mg/day, more than 190 mg/day, more than 200 mg/day, more than 225mg/day, more than 250 mg/day, more than 275 mg/day, more than 300mg/day, more than 325 mg/day, more than 350 mg/day, more than 375mg/day, more than 400 mg/day, more than 425 mg/day, more than 450mg/day, more than 475 mg/day, or more than 500 mg/day. In certainembodiments, the HDAC inhibitor is administered at a dose of more than 1mg/day and less than 500 mg/day. In some embodiments, the HDAC inhibitoris administered at a dose of more than 20 mg/day and less than 80mg/day. In certain embodiments, the HDAC inhibitor is administered oncea day (q.d.), twice a day (b.id.), or thrice a day (t.i.d.). In someembodiments, the HDAC inhibitor is administered daily, once a week,twice a week, three times a week, four times a week, or five times aweek.

In some embodiments, the HDAC inhibitor is CHR-3996. In certainembodiments, CHR-3996 is administered at a dose of 40 mg/day. In someembodiments, CHR-3996 is administered at a dose of about 1 mg/day, about2 mg/day, about 5 mg/day, about 10 mg/day, about 15 mg/day, about 20mg/day, about 25 mg/day, about 30 mg/day, about 35 mg/day, about 40mg/day, about 45 mg/day, about 50 mg/day, about 60 mg/day, about 70mg/day, about 80 mg/day, about 90 mg/day, or about 100 mg/day. Incertain embodiments, CHR-3996 is administered at a dose of less than 1mg/day, less than 2 mg/day, less than 5 mg/day, less than 10 mg/day,less than 15 mg/day, less than 20 mg/day, less than 25 mg/day, less than30 mg/day, less than 35 mg/day, less than 40 mg/day, less than 45mg/day, less than 50 mg/day, less than 60 mg/day, less than 70 mg/day,less than 80 mg/day, less than 90 mg/day, or less than 100 mg/day. Insome embodiments, CHR-3996 is administered at a dose of more than 1mg/day, more than 2 mg/day, more than 5 mg/day, more than 10 mg/day,more than 15 mg/day, more than 20 mg/day, more than 25 mg/day, more than30 mg/day, more than 35 mg/day, more than 40 mg/day, more than 45mg/day, more than 50 mg/day, more than 60 mg/day, more than 70 mg/day,more than 80 mg/day, more than 90 mg/day, or more than 100 mg/day. Incertain embodiments, CHR-3996 is administered at a dose of more than 30mg/day and less than 50 mg/day. In some embodiments, CHR-3996 isadministered at a dose of more than 20 mg/day and less than 80 mg/day.In certain embodiments, CHR-3996 is administered once a day (q.d.),twice a day (b.id.), or thrice a day (t.i.d.). In some embodiments,CHR-3996 is administered daily, once a week, twice a week, three times aweek, four times a week, or five times a week.

In some embodiments, a unit dose of a co-formulated HDAC inhibitor andantiviral agent comprises less than 100 mg of the HDAC inhibitor andless than 1000 mg of the antiviral agent. In certain embodiments, theunit dose comprises less than 50 mg of the HDAC inhibitor and less than500 mg of the antiviral agent. In other embodiments, the unit dosecomprises less than 80 mg of the HDAC inhibitor and less than 1500 mg ofthe antiviral agent. In some embodiments, the unit dose comprises lessthan 50 mg of the HDAC inhibitor CHR-3996 and less than 1000 mg ofvalganciclovir. In some embodiments, the unit dose comprises about 20 mgof the HDAC inhibitor CHR-3996 and about 450 mg of valganciclovir. Incertain embodiments, the unit dose comprises about 40 mg of the HDACinhibitor CHR-3996 and about 900 mg of valganciclovir. In someembodiments, the antiviral agent is formulated as controlled release.

In some embodiments, the HDAC inhibitor is ITF-2357. In certainembodiments, ITF-2357 is administered at a dose of 100 mg/day. In someembodiments, ITF-2357 is administered at a dose of about 1 mg/day, about2 mg/day, about 5 mg/day, about 10 mg/day, about 15 mg/day, about 20mg/day, about 25 mg/day, about 30 mg/day, about 35 mg/day, about 40mg/day, about 45 mg/day, about 50 mg/day, about 60 mg/day, about 70mg/day, about 80 mg/day, about 90 mg/day, about 100 mg/day, about 120mg/day, about 125 mg/day, about 140 mg/day, about 150 mg/day, about 160mg/day, about 175 mg/day, about 180 mg/day, about 200 mg/day, about 225mg/day, about 250 mg/day, about 275 mg/day, or about 300 mg/day. Incertain embodiments, ITF-2357 is administered at a dose of less than 1mg/day, less than 2 mg/day, less than 5 mg/day, less than 10 mg/day,less than 15 mg/day, less than 20 mg/day, less than 25 mg/day, less than30 mg/day, less than 35 mg/day, less than 40 mg/day, less than 45mg/day, less than 50 mg/day, less than 60 mg/day, less than 70 mg/day,less than 80 mg/day, less than 90 mg/day, less than 100 mg/day, lessthan 120 mg/day, less than 125 mg/day, less than 140 mg/day, less than150 mg/day, less than 160 mg/day, less than 175 mg/day, less than 180mg/day, less than 200 mg/day, less than 225 mg/day, less than 250mg/day, less than 275 mg/day, or less than 300 mg/day. In someembodiments, ITF-2357 is administered at a dose of more than 1 mg/day,more than 2 mg/day, more than 5 mg/day, more than 10 mg/day, more than15 mg/day, more than 20 mg/day, more than 25 mg/day, more than 30mg/day, more than 35 mg/day, more than 40 mg/day, more than 45 mg/day,more than 50 mg/day, more than 60 mg/day, more than 70 mg/day, more than80 mg/day, more than 90 mg/day, more than 100 mg/day, more than 120mg/day, more than 125 mg/day, more than 140 mg/day, more than 150mg/day, more than 160 mg/day, more than 175 mg/day, more than 180mg/day, more than 200 mg/day, more than 225 mg/day, more than 250mg/day, more than 275 mg/day, or more than 300 mg/day. In certainembodiments, ITF-2357 is administered at a dose of more than 80 mg/dayand less than 120 mg/day. In some embodiments, ITF-2357 is administeredat a dose of more than 40 mg/day and less than 120 mg/day. In certainembodiments, ITF-2357 is administered at a dose of more than 50 mg/dayand less than 240 mg/day. In some embodiments, ITF-2357 is administeredonce a day (q.d.), twice a day (b.id.), or thrice a day (t.i.d.). Insome embodiments, ITF-2357 is administered daily, once a week, twice aweek, three times a week, four times a week, or five times a week.

In some embodiments, the HDAC inhibitor is JNJ-16241199/R306465. Incertain embodiments, JNJ-16241199/R306465 is administered at a dose of100 mg/day. In some embodiments, JNJ-16241199/R306465 is administered ata dose of about 1 mg/day, about 2 mg/day, about 5 mg/day, about 10mg/day, about 15 mg/day, about 20 mg/day, about 25 mg/day, about 30mg/day, about 35 mg/day, about 40 mg/day, about 45 mg/day, about 50mg/day, about 60 mg/day, about 70 mg/day, about 80 mg/day, about 90mg/day, about 100 mg/day, about 120 mg/day, about 125 mg/day, about 140mg/day, about 150 mg/day, about 160 mg/day, about 175 mg/day, about 180mg/day, about 200 mg/day, about 225 mg/day, about 250 mg/day, about 275mg/day, or about 300 mg/day. In certain embodiments,JNJ-16241199/R306465 is administered at a dose of less than 1 mg/day,less than 2 mg/day, less than 5 mg/day, less than 10 mg/day, less than15 mg/day, less than 20 mg/day, less than 25 mg/day, less than 30mg/day, less than 35 mg/day, less than 40 mg/day, less than 45 mg/day,less than 50 mg/day, less than 60 mg/day, less than 70 mg/day, less than80 mg/day, less than 90 mg/day, less than 100 mg/day, less than 120mg/day, less than 125 mg/day, less than 140 mg/day, less than 150mg/day, less than 160 mg/day, less than 175 mg/day, less than 180mg/day, less than 200 mg/day, less than 225 mg/day, less than 250mg/day, less than 275 mg/day, or less than 300 mg/day. In someembodiments, JNJ-16241199/R306465 is administered at a dose of more than1 mg/day, more than 2 mg/day, more than 5 mg/day, more than 10 mg/day,more than 15 mg/day, more than 20 mg/day, more than 25 mg/day, more than30 mg/day, more than 35 mg/day, more than 40 mg/day, more than 45mg/day, more than 50 mg/day, more than 60 mg/day, more than 70 mg/day,more than 80 mg/day, more than 90 mg/day, more than 100 mg/day, morethan 120 mg/day, more than 125 mg/day, more than 140 mg/day, more than150 mg/day, more than 160 mg/day, more than 175 mg/day, more than 180mg/day, more than 200 mg/day, more than 225 mg/day, more than 250mg/day, more than 275 mg/day, or more than 300 mg/day. In certainembodiments, JNJ-16241199/R306465 is administered at a dose of more than80 mg/day and less than 120 mg/day. In some embodiments,JNJ-16241199/R306465 is administered at a dose of more than 40 mg/dayand less than 120 mg/day. In certain embodiments, JNJ-16241199/R306465is administered at a dose of more than 50 mg/day and less than 240mg/day. In some embodiments, JNJ-16241199/R306465 is administered once aday (q.d.), twice a day (b.id.), or thrice a day (t.i.d.). In someembodiments, JNJ-16241199/R306465 is administered daily, once a week,twice a week, three times a week, four times a week, or five times aweek.

In some embodiments, the HDAC inhibitor is JNJ-26481585. In certainembodiments, JNJ-26481585 is administered at a dose of 10 mg/day. Insome embodiments, JNJ-26481585 is administered at a dose of about 0.1mg/day, about 0.2 mg/day, about 0.5 mg/day, about 1 mg/day, about 2mg/day, about 5 mg/day, about 10 mg/day, about 15 mg/day, about 20mg/day, about 25 mg/day, about 30 mg/day, about 35 mg/day, about 40mg/day, about 45 mg/day, about 50 mg/day, about 60 mg/day, about 70mg/day, about 80 mg/day, about 90 mg/day, or about 100 mg/day. Incertain embodiments, JNJ-26481585 is administered at a dose of less than0.1 mg/day, less than 0.2 mg/day, less than 0.5 mg/day, less than 1mg/day, less than 2 mg/day, less than 5 mg/day, less than 10 mg/day,less than 15 mg/day, less than 20 mg/day, less than 25 mg/day, less than30 mg/day, less than 35 mg/day, less than 40 mg/day, less than 45mg/day, less than 50 mg/day, less than 60 mg/day, less than 70 mg/day,less than 80 mg/day, less than 90 mg/day, or less than 100 mg/day. Insome embodiments, JNJ-26481585 is administered at a dose of more than0.1 mg/day, more than 0.2 mg/day, more than 0.5 mg/day, more than 1mg/day, more than 2 mg/day, more than 5 mg/day, more than 10 mg/day,more than 15 mg/day, more than 20 mg/day, more than 25 mg/day, more than30 mg/day, more than 35 mg/day, more than 40 mg/day, more than 45mg/day, more than 50 mg/day, more than 60 mg/day, more than 70 mg/day,more than 80 mg/day, more than 90 mg/day, or more than 100 mg/day. Incertain embodiments, JNJ-26481585 is administered at a dose of more than2 mg/day and less than 20 mg/day. In some embodiments, JNJ-26481585 isadministered at a dose of more than 5 mg/day and less than 30 mg/day. Incertain embodiments, JNJ-26481585 is administered once a day (q.d.),twice a day (b.id.), or thrice a day (t.i.d.). In some embodiments,JNJ-26481585 is administered daily, once a week, twice a week, threetimes a week, four times a week, or five times a week.

In some embodiments, the HDAC inhibitor is MGCD103. In certainembodiments, MGCD103 is administered at a dose of 45 mg/m²/day. In someembodiments, MGCD103 is administered at a dose of about 1 mg/m²/day,about 2 mg/m²/day, about 5 mg/m²/day, about 10 mg/m²/day, about 15mg/m²/day, about 20 mg/m²/day, about 25 mg/m²/day, about 30 mg/m²/day,about 35 mg/m²/day, about 40 mg/m2/day, about 45 mg/m²/day, about 50mg/m²/day, about 60 mg/m²/day, about 70 mg/m²/day, about 80 mg/m²/day,about 90 mg/m²/day, or about 100 mg/m²/day. In certain embodiments,MGCD103 is administered at a dose of less than 1 mg/m²/day, less than 2mg/m²/day, less than 5 mg/m²/day, less than 10 mg/m²/day, less than 15mg/m²/day, less than 20 mg/m²/day, less than 25 mg/m²/day, less than 30mg/m²/day, less than 35 mg/m²/day, less than 40 mg/m²/day, less than 45mg/m²/day, less than 50 mg/m²/day, less than 60 mg/m²/day, less than 70mg/m²/day, less than 80 mg/m²/day, less than 90 mg/m²/day, or less than100 mg/m²/day. In some embodiments, MGCD103 is administered at a dose ofmore than 1 mg/m²/day, more than 2 mg/m²/day, more than 5 mg/m²/day,more than 10 mg/m²/day, more than 15 mg/m²/day, more than 20 mg/m²/day,more than 25 mg/m²/day, more than 30 mg/m²/day, more than 35 mg/m²/day,more than 40 mg/m²/day, more than 45 mg/m²/day, more than 50 mg/m²/day,more than 60 mg/m²/day, more than 70 mg/m²/day, more than 80 mg/m²/day,more than 90 mg/m²/day, or more than 100 mg/m²/day. In certainembodiments, MGCD103 is administered at a dose of more than 30 mg/m²/dayand less than 80 mg/m²/day. In some embodiments, MGCD103 is administeredat a dose of more than 45 mg/m²/day and less than 60 mg/m²/day. Incertain embodiments, MGCD103 is administered once a day (q.d.), twice aday (b.id.), or thrice a day (t.i.d.). In some embodiments, MGCD103 isadministered daily, once a week, twice a week, three times a week, fourtimes a week, or five times a week.

In some embodiments, the HDAC inhibitor is MS-275. In certainembodiments, MS-275 is administered at a dose of 4 mg/m²/day. In someembodiments, MS-275 is administered at a dose of about 0.1 mg/m²/day, ofabout 0.2 mg/m²/day, of about 0.5 mg/m²/day, of about 1 mg/m²/day, ofabout 2 mg/m²/day, of about 3 mg/m²/day, of about 4 mg/m²/day, about 5mg/m²/day, of about 6 mg/m²/day, of about 7 mg/m²/day, of about 8mg/m²/day, of about 9 mg/m²/day, about 10 mg/m²/day, about 15 mg/m²/day,about 20 mg/m²/day, about 25 mg/m²/day, about 30 mg/m²/day, about 35mg/m²/day, about 40 mg/m2/day, about 45 mg/m²/day, about 50 mg/m²/day,about 60 mg/m²/day, about 70 mg/m²/day, about 80 mg/m²/day, about 90mg/m²/day, or about 100 mg/m²/day. In certain embodiments, MS-275 isadministered at a dose of less than 0.1 mg/m²/day, of less than 0.2mg/m²/day, of less than 0.5 mg/m²/day, of less than 1 mg/m²/day, of lessthan 2 mg/m²/day, of less than 3 mg/m²/day, of less than 4 mg/m²/day,less than 5 mg/m²/day, of less than 6 mg/m²/day, of less than 7mg/m²/day, of less than 8 mg/m²/day, of less than 9 mg/m²/day, less than10 mg/m²/day, less than 15 mg/m²/day, less than 20 mg/m²/day, less than25 mg/m²/day, less than 30 mg/m²/day, less than 35 mg/m²/day, less than40 mg/m2/day, less than 45 mg/m²/day, less than 50 mg/m²/day, less than60 mg/m²/day, less than 70 mg/m²/day, less than 80 mg/m²/day, less than90 mg/m²/day, or less than 100 mg/m²/day. In some embodiments, MS-275 isadministered at a dose of more than 0.1 mg/m²/day, of more than 0.2mg/m²/day, of more than 0.5 mg/m²/day, of more than 1 mg/m²/day, of morethan 2 mg/m²/day, of more than 3 mg/m²/day, of more than 4 mg/m²/day,more than 5 mg/m²/day, of more than 6 mg/m²/day, of more than 7mg/m²/day, of more than 8 mg/m²/day, of more than 9 mg/m²/day, more than10 mg/m²/day, more than 15 mg/m²/day, more than 20 mg/m²/day, more than25 mg/m²/day, more than 30 mg/m²/day, more than 35 mg/m²/day, more than40 mg/m2/day, more than 45 mg/m²/day, more than 50 mg/m²/day, more than60 mg/m²/day, more than 70 mg/m²/day, more than 80 mg/m²/day, more than90 mg/m²/day, or more than 100 mg/m²/day. In certain embodiments, MS-275is administered at a dose of more than 2 mg/m²/day and less than 10mg/m²/day. In some embodiments, MS-275 is administered at a dose of morethan 2 mg/m²/day and less than 40 mg/m²/day. In certain embodiments,MS-275 is administered at a dose of more than 2 mg/m²/day and less than6 mg/m²/day. In some embodiments, MS-275 is administered at a dose ofmore than 6 mg/m²/day and less than 8 mg/m²/day. In certain embodiments,MS-275 is administered once a day (q.d.), twice a day (b.id.), or thricea day (t.i.d.). In some embodiments, MS-275 is administered daily, oncea week, twice a week, three times a week, four times a week, or fivetimes a week.

In some embodiments, the HDAC inhibitor is SB939. In certainembodiments, SB939 is administered at a dose of 60 mg/day. In someembodiments, SB939 is administered at a dose of about 1 mg/day, about 2mg/day, about 5 mg/day, about 10 mg/day, about 15 mg/day, about 20mg/day, about 25 mg/day, about 30 mg/day, about 35 mg/day, about 40mg/day, about 45 mg/day, about 50 mg/day, about 60 mg/day, about 70mg/day, about 80 mg/day, about 90 mg/day, about 100 mg/day, about 120mg/day, about 125 mg/day, about 140 mg/day, about 150 mg/day, about 160mg/day, about 175 mg/day, about 180 mg/day, or about 200 mg/day. Incertain embodiments, SB939 is administered at a dose of less than 1mg/day, less than 2 mg/day, less than 5 mg/day, less than 10 mg/day,less than 15 mg/day, less than 20 mg/day, less than 25 mg/day, less than30 mg/day, less than 35 mg/day, less than 40 mg/day, less than 45mg/day, less than 50 mg/day, less than 60 mg/day, less than 70 mg/day,less than 80 mg/day, less than 90 mg/day, less than 100 mg/day, lessthan 120 mg/day, less than 125 mg/day, less than 140 mg/day, less than150 mg/day, less than 160 mg/day, less than 175 mg/day, less than 180mg/day, or less than 200 mg/day. In some embodiments, SB939 isadministered at a dose of more than 1 mg/day, more than 2 mg/day, morethan 5 mg/day, more than 10 mg/day, more than 15 mg/day, more than 20mg/day, more than 25 mg/day, more than 30 mg/day, more than 35 mg/day,more than 40 mg/day, more than 45 mg/day, more than 50 mg/day, more than60 mg/day, more than 70 mg/day, more than 80 mg/day, more than 90mg/day, more than 100 mg/day, more than 120 mg/day, more than 125mg/day, more than 140 mg/day, more than 150 mg/day, more than 160mg/day, more than 175 mg/day, more than 180 mg/day, or more than 200mg/day. In certain embodiments, SB939 is administered at a dose of morethan 30 mg/day and less than 70 mg/day. In some embodiments, SB939 isadministered at a dose of more than 10 mg/day and less than 90 mg/day.In certain embodiments, SB939 is administered once a day (q.d.), twice aday (b.id.), or thrice a day (t.i.d.). In some embodiments, SB939 isadministered daily, once a week, twice a week, three times a week, fourtimes a week, or five times a week.

In some embodiments, the HDAC inhibitor is romidepsin. In certainembodiments, romidepsin is administered at a dose of 14 mg/m²/day. Insome embodiments, romidepsin is administered at a dose of about 0.1mg/m²/day, of about 0.2 mg/m²/day, of about 0.5 mg/m²/day, of about 1mg/m²/day, of about 2 mg/m²/day, of about 3 mg/m²/day, of about 4mg/m²/day, about 5 mg/m²/day, of about 6 mg/m²/day, of about 7mg/m²/day, of about 8 mg/m²/day, of about 9 mg/m²/day, about 10mg/m²/day, about 11 mg/m²/day, about 12 mg/m²/day, about 13 mg/m²/day,about 14 mg/m²/day, about 15 mg/m²/day, about 16 mg/m²/day, about 17mg/m²/day, about 18 mg/m²/day, about 19 mg/m²/day, about 20 mg/m²/day,about 25 mg/m²/day, about 30 mg/m²/day, about 35 mg/m²/day, about 40mg/m2/day, about 45 mg/m²/day, about 50 mg/m²/day, about 60 mg/m²/day,about 70 mg/m²/day, about 80 mg/m²/day, about 90 mg/m²/day, or about 100mg/m²/day. In certain embodiments, romidepsin is administered at a doseof less than 0.1 mg/m²/day, of less than 0.2 mg/m²/day, of less than 0.5mg/m²/day, of less than 1 mg/m²/day, of less than 2 mg/m²/day, of lessthan 3 mg/m²/day, of less than 4 mg/m²/day, less than 5 mg/m²/day, ofless than 6 mg/m²/day, of less than 7 mg/m²/day, of less than 8mg/m²/day, of less than 9 mg/m²/day, less than 10 mg/m²/day, less than11 mg/m²/day, less than 12 mg/m²/day, less than 13 mg/m²/day, less than14 mg/m²/day, less than 15 mg/m²/day, less than 16 mg/m²/day, less than17 mg/m²/day, less than 18 mg/m²/day, less than 19 mg/m²/day, less than20 mg/m²/day, less than 25 mg/m²/day, less than 30 mg/m²/day, less than35 mg/m²/day, less than 40 mg/m2/day, less than 45 mg/m²/day, less than50 mg/m²/day, less than 60 mg/m²/day, less than 70 mg/m²/day, less than80 mg/m²/day, less than 90 mg/m²/day, or less than 100 mg/m²/day. Insome embodiments, romidepsin is administered at a dose of more than 0.1mg/m²/day, of more than 0.2 mg/m²/day, of more than 0.5 mg/m²/day, ofmore than 1 mg/m²/day, of more than 2 mg/m²/day, of more than 3mg/m²/day, of more than 4 mg/m²/day, more than 5 mg/m²/day, of more than6 mg/m²/day, of more than 7 mg/m²/day, of more than 8 mg/m²/day, of morethan 9 mg/m²/day, more than 10 mg/m²/day, more than 11 mg/m²/day, morethan 12 mg/m²/day, more than 13 mg/m²/day, more than 14 mg/m²/day, morethan 15 mg/m²/day, more than 16 mg/m²/day, more than 17 mg/m²/day, morethan 18 mg/m²/day, more than 19 mg/m²/day, more than 20 mg/m²/day, morethan 25 mg/m²/day, more than 30 mg/m²/day, more than 35 mg/m²/day, morethan 40 mg/m2/day, more than 45 mg/m²/day, more than 50 mg/m²/day, morethan 60 mg/m²/day, more than 70 mg/m²/day, more than 80 mg/m²/day, morethan 90 mg/m²/day, or more than 100 mg/m²/day. In certain embodiments,romidepsin is administered at a dose of more than 13 mg/m²/day and lessthan 18 mg/m²/day. In some embodiments, romidepsin is administered at adose of more than 10 mg/m²/day and less than 20 mg/m²/day. In certainembodiments, romidepsin is administered once a day (q.d.), twice a day(b.id.), or thrice a day (t.i.d.). In some embodiments, romidepsin isadministered daily, once a week, twice a week, three times a week, fourtimes a week, or five times a week.

In some embodiments, the HDAC inhibitor is LBH589. In certainembodiments, LBH589 is administered at a dose of 20 mg/day. In someembodiments, LBH589 is administered at a dose of about 1 mg/day, about 2mg/day, about 5 mg/day, about 10 mg/day, about 15 mg/day, about 20mg/day, about 25 mg/day, about 30 mg/day, about 35 mg/day, about 40mg/day, about 45 mg/day, about 50 mg/day, about 60 mg/day, about 70mg/day, about 80 mg/day, about 90 mg/day, or about 100 mg/day. Incertain embodiments, LBH589 is administered at a dose of less than 1mg/day, less than 2 mg/day, less than 5 mg/day, less than 10 mg/day,less than 15 mg/day, less than 20 mg/day, less than 25 mg/day, less than30 mg/day, less than 35 mg/day, less than 40 mg/day, less than 45mg/day, less than 50 mg/day, less than 60 mg/day, less than 70 mg/day,less than 80 mg/day, less than 90 mg/day, or less than 100 mg/day. Insome embodiments, LBH589 is administered at a dose of more than 1mg/day, more than 2 mg/day, more than 5 mg/day, more than 10 mg/day,more than 15 mg/day, more than 20 mg/day, more than 25 mg/day, more than30 mg/day, more than 35 mg/day, more than 40 mg/day, more than 45mg/day, more than 50 mg/day, more than 60 mg/day, more than 70 mg/day,more than 80 mg/day, more than 90 mg/day, or more than 100 mg/day. Incertain embodiments, LBH589 is administered at a dose of more than 10mg/day and less than 20 mg/day. In some embodiments, LBH589 isadministered at a dose of more than 5 mg/day and less than 30 mg/day. Incertain embodiments, LBH589 is administered once a day (q.d.), twice aday (b.id.), or thrice a day (t.i.d.). In some embodiments, LBH589 isadministered daily, once a week, twice a week, three times a week, fourtimes a week, or five times a week.

In some embodiments, the HDAC inhibitor is PXD101. In certainembodiments, PXD101 is administered at a dose of 1000 mg/m²/day. In someembodiments, PXD101 is administered at a dose of about 10 mg/m²/day,about 15 mg/m²/day, about 20 mg/m²/day, about 50 mg/m²/day, about 75mg/m²/day, about 100 mg/m²/day, about 150 mg/m²/day, about 200mg/m²/day, about 300 mg/m²/day, about 400 mg/m²/day, about 500mg/m²/day, about 600 mg/m²/day, about 700 mg/m²/day, about 800mg/m²/day, about 900 mg/m²/day, about 1000 mg/m²/day, about 1100mg/m²/day, about 1200 mg/m²/day, about 1300 mg/m²/day, about 1400mg/m²/day, about 1500 mg/m²/day, about 1750 mg/m²/day, or about 2000mg/m²/day. In certain embodiments, PXD101 is administered at a dose ofless than 10 mg/m²/day, less than 15 mg/m²/day, less than 20 mg/m²/day,less than 50 mg/m²/day, less than 75 mg/m²/day, less than 100 mg/m²/day,less than 150 mg/m²/day, less than 200 mg/m²/day, less than 300mg/m²/day, less than 400 mg/m²/day, less than 500 mg/m²/day, less than600 mg/m²/day, less than 700 mg/m²/day, less than 800 mg/m²/day, lessthan 900 mg/m²/day, less than 1000 mg/m²/day, less than 1100 mg/m²/day,less than 1200 mg/m²/day, less than 1300 mg/m²/day, less than 1400mg/m²/day, less than 1500 mg/m²/day, less than 1750 mg/m²/day, or lessthan 2000 mg/m²/day. In some embodiments, PXD101 is administered at adose of more than 10 mg/m²/day, more than 15 mg/m²/day, more than 20mg/m²/day, more than 50 mg/m²/day, more than 75 mg/m²/day, more than 100mg/m²/day, more than 150 mg/m²/day, more than 200 mg/m²/day, more than300 mg/m²/day, more than 400 mg/m²/day, more than 500 mg/m²/day, morethan 600 mg/m²/day, more than 700 mg/m²/day, more than 800 mg/m²/day,more than 900 mg/m²/day, more than 1000 mg/m²/day, more than 1100mg/m²/day, more than 1200 mg/m²/day, more than 1300 mg/m²/day, more than1400 mg/m²/day, more than 1500 mg/m²/day, more than 1750 mg/m²/day, ormore than 2000 mg/m²/day. In certain embodiments, PXD101 is administeredat a dose of more than 600 mg/m²/day and less than 1000 mg/m²/day. Insome embodiments, PXD101 is administered at a dose of more than 15mg/m²/day and less than 1000 mg/m²/day. In certain embodiments, PXD101is administered once a day (q.d.), twice a day (b.id.), or thrice a day(t.i.d.). In some embodiments, PXD101 is administered daily, once aweek, twice a week, three time a week, four times a week, or five timesa week.

In some embodiments, the HDAC inhibitor is vorinostat. In certainembodiments, vorinostat is administered at a dose of 400 mg/day. In someembodiments, vorinostat is administered at a dose of about 10 mg/day,about 20 mg/day, about 30 mg/day, about 40 mg/day, about 50 mg/day,about 60 mg/day, about 70 mg/day, about 80 mg/day, about 90 mg/day,about 100 mg/day, about 125 mg/day, about 150 mg/day, about 175 mg/day,about 200 mg/day, about 225 mg/day, about 250 mg/day, about 275 mg/day,about 300 mg/day, about 325 mg/day, about 350 mg/day, about 375 mg/day,about 400 mg/day, about 450 mg/day, about 500 mg/day, about 550 mg/day,about 600 mg/day, about 650 mg/day, about 700 mg/day, about 750 mg/day,about 800 mg/day, about 900 mg/day, or about 1000 mg/day. In certainembodiments, vorinostat is administered at a dose of less than 10mg/day, less than 20 mg/day, less than 30 mg/day, less than 40 mg/day,less than 50 mg/day, less than 60 mg/day, less than 70 mg/day, less than80 mg/day, less than 90 mg/day, less than 100 mg/day, less than 125mg/day, less than 150 mg/day, less than 175 mg/day, less than 200mg/day, less than 225 mg/day, less than 250 mg/day, less than 275mg/day, less than 300 mg/day, less than 325 mg/day, less than 350mg/day, less than 375 mg/day, less than 400 mg/day, less than 450mg/day, less than 500 mg/day, less than 550 mg/day, less than 600mg/day, less than 650 mg/day, less than 700 mg/day, less than 750mg/day, less than 800 mg/day, less than 900 mg/day, or less than 1000mg/day. In some embodiments, vorinostat is administered at a dose ofmore than 10 mg/day, more than 20 mg/day, more than 30 mg/day, more than40 mg/day, more than 50 mg/day, more than 60 mg/day, more than 70mg/day, more than 80 mg/day, more than 90 mg/day, more than 100 mg/day,more than 125 mg/day, more than 150 mg/day, more than 175 mg/day, morethan 200 mg/day, more than 225 mg/day, more than 250 mg/day, more than275 mg/day, more than 300 mg/day, more than 325 mg/day, more than 350mg/day, more than 375 mg/day, more than 400 mg/day, more than 450mg/day, more than 500 mg/day, more than 550 mg/day, more than 600mg/day, more than 650 mg/day, more than 700 mg/day, more than 750mg/day, more than 800 mg/day, more than 900 mg/day, or more than 1000mg/day. In certain embodiments, vorinostat is administered at a dose ofmore than 100 mg/day and less than 400 mg/day. In some embodiments,vorinostat is administered at a dose of more than 100 mg/day and lessthan 500 mg/day. In certain embodiments, vorinostat is administered oncea day (q.d.), twice a day (b.id.), or thrice a day (t.i.d.). In someembodiments, vorinostat is administered daily, once a week, twice aweek, three times a week, four times a week, or five times a week.

In some embodiments, an HDAC inhibitor inhibits the growth ofvirus-positive cells. In certain embodiments, the HDAC inhibitorinhibits the growths of EBV-positive lymphoma cells. In someembodiments, the HDAC inhibitor has growth inhibitory activity at aconcentration of about 100 μM, about 90 μM, about 80 μM, about 75 μM,about 70 μM, about 60 μM, about 50 μM, about 40 μM, about 30 μM, about25 μM, about 20 μM, about 10 μM, about 5 μM, about 2 μM, about 1 μM,about 900 nM, about 800 nM, about 700 nM, about 600 nM, about 500 nM,about 400 nM, about 300 nM, about 200 nM, about 100 nM, about 75 nM,about 50 nM, about 20 nM, or about 10 nM. In certain embodiments, theHDAC inhibitor has growth inhibitory activity at a concentration of lessthan 100 μM, less than 90 μM, less than 80 μM, less than 75 μM, lessthan 70 μM, less than 60 μM, less than 50 μM, less than 40 μM, less than30 μM, less than 25 μM, less than 20 μM, less than 10 μM, less than 5μM, less than 2 μM, less than 1 μM, less than 900 nM, less than 800 nM,less than 700 nM, less than 600 nM, less than 500 nM, less than 400 nM,less than 300 nM, less than 200 nM, less than 100 nM, less than 75 nM,less than 50 nM, less than 20 nM, or less than 10 nM. In someembodiments, the HDAC inhibitor has growth inhibitory activity at aconcentration of more than 100 μM, more than 90 μM, more than 80 μM,more than 75 μM, more than 70 μM, more than 60 μM, more than 50 μM, morethan 40 μM, more than 30 μM, more than 25 μM, more than 20 μM, more than10 μM, more than 5 μM, more than 2 μM, more than 1 μM, more than 900 nM,more than 800 nM, more than 700 nM, more than 600 nM, more than 500 nM,more than 400 nM, more than 300 nM, more than 200 nM, more than 100 nM,more than 75 nM, more than 50 nM, more than 20 nM, or more than 10 nM.In certain embodiments, the HDAC inhibitor has growth inhibitoryactivity at more than 50 nM and less than 100 nM. In some embodiments,the HDAC inhibitor has growth inhibitory activity at more than 200 nMand less than 500 nM. In certain embodiments, the HDAC inhibitor hasgrowth inhibitory activity at more than 100 nM and less than 200 nM.

Induced Genes Including Viral-Associated Genes

Inducing agents (agents that induce expression) may act directly on theviral genome or indirectly through a cellular factor required for viralexpression. For example, viral gene expression can be regulated throughthe regulation of the expression of viral transcription factors such asZTA, RTA, tat, and tax, cellular transcription factors such as AP-1,AP-2, Sp 1, NF-icB, and other transcriptional activators and/orrepressors (factors), co-activators and co-repressors, histoneacetylators and deacetylators, DNA methylases and demethylases,oncogenes or proto-oncogenes, or protein kinase C. These proteins act toregulate and thereby control expression of specific viral and/or othercellular genetic elements. According to the methods of the invention,control over their expression can lead to control over the infection.Other gene products, both viral and cellular in origin, whose expressioncan be regulated with inducing agents include proteases, polymerases,reverse transcriptases, cell-surface receptors, major histocompatibilityantigens, growth factors, and combination of these products.

Additional genes whose expression or transcriptional regulation arealtered in the presence of butyric acid include the oncogenes myc, ras,myb, abl and src. The activities of these gene products, as well as theactivities of other oncogenes, are described in Slamon, J. D., et al.1984 Science 224:256-62. Anti-proliferative activity also includes theability to repress tumor angiogenesis through the blockade ofangiogenesis factor activity, production or release, transcriptionalregulation, or the ability to modulate transcription of genes underangiogenesis or growth factor or holinonal control. Either would be aneffective therapy, particularly against both prostatic neoplasia andbreast carcinomas. Further activities that effect transcription and/orcellular differentiation include increased intracellular cAMP levels,inhibition of histone acetylation, and inhibition of genomicmethylation. Each of these activities is directly related to geneexpression, and increased expression can sensitize infected cells to aspecific anti-viral agent.

In some embodiments, inducing agents include arginine butyrate and/orother histone deacetylase inhibitors. Arginine butyrate induces EBV-TKactivity in EBV-immortalized B-cells and patient-derived tumor cells. Aslatently-infected B-cells do not express TK, exposure of these cells toagents like arginine butyrate and/or other HDAC inhibitors results in ainduction of lytic replication and TK expression. This expression of aviral gene can be used as a point for attack by anti-viral agents,allowing for treatment of latent infections.

In other embodiments, inducing agents include HDAC inhibitors thatinduce EBV-PK activity (also known BGLF4) in EBV infected tumors.Expression of EBV-PK/BGLF4 sensitizes a cell to an antiviral agent. Incertain instances, HDAC inhibitors induce EBV-PK. In some instances,HDAC inhibitors induce EBV-TK and/or EBV-PK.

Preliminary in vitro studies according to the invention demonstrate thatinduction of EBV-TK activity in EBV-immortalized B-cells andpatient-derived tumor cells using these drugs is possible, and thatthese previously resistant cells are rendered susceptible to ganciclovirtherapy. Treatment of patients with viral-associated tumors such as EBVwith inducing agents such as arginine butyrate, to induce the expressionof EBV-TK/EBV-PK, and GCV, to eliminate EBV-TK/EBV-PK expressing tumorcells, is an effective, non-toxic therapy. This therapeutic regimen doesnot depend on the associated viral genome being the cause of the tumor.Without wishing to be bound by theory, it is believed that just thepresence of the EBV genome in latent form would make the tumorsusceptible to this combination protocol.

Butyrate-associated induction of genes has been characterized forvarious cell types, and the genes are consistently in the class ofdifferentiation markers of a cell. For example, in colon cancer celllines, morphologic changes observed in the presence of butyratecorrelate with increased expression of alkaline phosphatase, plasminogenactivator, and CEA, all markers of differentiation. Hepatoma cell linesincrease expression of alpha fetoprotein. Breast cancer cell linesexpress milk-related glycoproteins, epithelial membrane antigens, andincreased lipid deposition. Sodium butyrate can also induce expressionof cellular proteins associated with converting basal keratinacytes intocommitted epithelial cells.

Alteration of expression of certain transcription factors may affectregulation of gene expression and regulation of the cell cycle. In thebreast cancer cell line MCF-7, butyrate induces a block in cellularproliferation that is associated with decreased expression of estrogenand prolactin hormone receptor mRNA expression, thus blocking thepotential growth stimulation by estrogen and prolactin. These effectsare associated with increased expression of the EGF receptor. Butyratealso has been shown to induce down-regulation of c-myc and p53 mRNA andto up-regulate expression of the c-fos transcription factor. In mousefibroblasts, butyrate will block the cell cycle in the G₁ phase. Whenthese cells are stimulated to proliferate with serum, TPA, or insulin,the immediate-early response transcription factors c-myc and c-jun areunregulated. However, the late G₁ phase downstream gene marker cdc-2mRNA is not expressed, and cells are prevented from entering S phase.

The particular combination of inducing agent with anti-viral agent thatis most effective against a specific disorder can be determined by oneof ordinary skill in the art from empirical testing and, preferably,from a knowledge of each agent's mechanism of action. Three suchexamples are as follows. First, many of the RNA viruses such as HIV andother retroviruses require a reverse transcriptase to transcribe theirgenome into DNA. A few of the agents that induce expression or activityof retroviruses and their encoded genes, such as, for example, reversetranscriptase, are known to those of ordinary skill in the art.Anti-viral agents such as nucleoside analogs can be administered to thepatient. Those substrate analogs will be specifically recognized by thereverse transcriptase that, when incorporated into the infected-cellgenome, prevent viral replication and may also result in cell death.Second, many viruses require an active protease to assemble viruscapsids to be packaged with viral genome. Protease inhibitors orproteases that alter cleavage patterns so that packaging cannot occurcan be specifically targeted with an anti-viral agent that comprises anamino acid analog or toxic conjugate. Third, arginine butyrate andisobutyramide enhance expression of viral thymidine kinase and otherviral protein kinases in EBV-infected lymphocytes. Ganciclovir orfamcyclovir, in the presence of the viral thymidine kinase or otherviral kinases, destroys the infected cell. Treatment of infected cellswith both agents, according to the invention, will selectively destroyEBV virus-infected cells. In another aspect, of infected cells with bothagents, according to the invention, will selectively disable or disruptthe viral activity within the cells in vivo.

In some embodiments, an inducing agent induces viral gene expression bymore than 4 fold after 24 h of treatment. In certain embodiments, anHDAC inhibitor induces TK or EBV-PK expression by more than 4 fold after24 h of treatment. In some embodiments, an HDAC inhibitor induces viralgene expression after about 48 h, about 36 h, about 24 h, about 18 h,about 12 h, about 8 h, about 6 h, about 4 h, about 3 h, about 2 h, about1 h, or about 30 minutes. In certain embodiments, an HDAC inhibitorinduces viral gene expression in less than 48 h, less than 36 h, lessthan 24 h, less than 18 h, less than 12 h, less than 8 h, less than 6 h,less than 4 h, less than 3 h, less than 2 h, less than 1 h, or less than30 minutes. In some embodiments, an HDAC inhibitor induces viral geneexpression in more than 48 h, more than 36 h, more than 24 h, more than18 h, more than 12 h, more than 8 h, more than 6 h, more than 4 h, morethan 3 h, more than 2 h, more than 1 h, or more than 30 minutes. Incertain embodiments, an HDAC inhibitor induces viral gene expressionafter more than 30 minutes and less than 24 h.

In certain embodiments, an inducing agent is capable of inducing geneexpression at a concentration of less than 500 nM. In some embodiments,the inducing agent is an HDAC inhibitor. In certain embodiments, theinducing agent is capable of inducing TK or EBV-PK expression. Incertain embodiments, an inducing agent is capable of inducing geneexpression at a concentration of about 100 μM, about 90 μM, about 80 μM,about 75 μM, about 70 μM, about 60 μM, about 50 μM, about 40 μM, about30 μM, about 25 μM, about 20 μM, about 10 μM, about 5 μM, about 2 μM,about 1 μM, about 900 nM, about 800 nM, about 700 nM, about 600 nM,about 500 nM, about 400 nM, about 300 nM, about 200 nM, about 100 nM,about 75 nM, about 50 nM, about 20 nM, or about 10 nM. In someembodiments, an inducing agent is capable of inducing gene expression ata concentration of less than 100 μM, less than 90 μM, less than 80 μM,less than 75 μM, less than 70 μM, less than 60 μM, less than 50 μM, lessthan 40 μM, less than 30 μM, less than 25 μM, less than 20 μM, less than10 μM, less than 5 μM, less than 2 μM, less than 1 μM, less than 900 nM,less than 800 nM, less than 700 nM, less than 600 nM, less than 500 nM,less than 400 nM, less than 300 nM, less than 200 nM, less than 100 nM,less than 75 nM, less than 50 nM, less than 20 nM, or less than 10 nM.In certain embodiments, an inducing agent is capable of inducing geneexpression at a concentration of more than 100 μM, more than 90 μM, morethan 80 μM, more than 75 μM, more than 70 μM, more than 60 μM, more than50 μM, more than 40 μM, more than 30 μM, more than 25 μM, more than 20μM, more than 10 μM, more than 5 μM, more than 2 μM, more than 1 μM,more than 900 nM, more than 800 nM, more than 700 nM, more than 600 nM,more than 500 nM, more than 400 nM, more than 300 nM, more than 200 nM,more than 100 nM, more than 75 nM, more than 50 nM, more than 20 nM, ormore than 10 nM. In some embodiments, an inducing agent is capable ofinducing gene expression at a concentration more than 50 nM and lessthan 100 nM. In certain embodiments, an inducing agent is capable ofinducing gene expression at a concentration of more than 200 nM and lessthan 500 nM. In some embodiments, an inducing agent is capable ofinducing gene expression at more than 100 nM and less than 200 nM

In some embodiments, an HDAC inhibitor induces viral gene expressionafter more than 1 h and less than 6 h. In certain embodiments, an HDACinhibitor induces viral gene expression about 2 fold, about 3 fold,about 4 fold, about 5 fold, about 6 fold, about 7 fold, about 8 fold,about 9 fold, about 10 fold, about 12 fold, about 15 fold, about 20fold, about 25 fold, about 30 fold, about 35 fold, about 40 fold, about45 fold, or about 50 fold. In some embodiments, an HDAC inhibitorinduces viral gene expression less than 2 fold, less than 3 fold, lessthan 4 fold, less than 5 fold, less than 6 fold, less than 7 fold, lessthan 8 fold, less than 9 fold, less than 10 fold, less than 12 fold,less than 15 fold, less than 20 fold, less than 25 fold, less than 30fold, less than 35 fold, less than 40 fold, less than 45 fold, or lessthan 50 fold. In certain embodiments, an HDAC inhibitor induces viralgene expression more than 2 fold, more than 3 fold, more than 4 fold,more than 5 fold, more than 6 fold, more than 7 fold, more than 8 fold,more than 9 fold, more than 10 fold, more than 12 fold, more than 15fold, more than 20 fold, more than 25 fold, more than 30 fold, more than35 fold, more than 40 fold, more than 45 fold, or more than 50 fold. Insome embodiments, an HDAC inhibitor induces viral gene expression morethan 2 fold and less than 50 fold. In certain embodiments, an HDACinhibitor induces viral gene expression more than 5 fold and less than40 fold.

Antiviral Agents

Anti-viral agents that can be used in the compositions and methods ofthe provided invention can include, for example, substrates andsubstrate analogs, inhibitors and other agents that severely impair,debilitate or otherwise destroy virus-infected cells. Substrate analogsinclude amino acid and nucleoside analogs. Substrates can be conjugatedwith toxins or other viricidal substances. Inhibitors include integraseinhibitors, protease inhibitors, polymerase inhibitors and transcriptaseinhibitors such as reverse transcriptase inhibitors.

Antiviral agents that can be used in the compositions and methods of theprovided invention can include, for example, ganciclovir,valganciclovir, oseltamivir (Tamiflu™), zanamivir (Relenza™), abacavir,aciclovir, acyclovir, adefovir, amantadine, amprenavir, ampligen,arbidol, atazanavir, atripla, boceprevir, cidofovir, combivir,darunavir, delavirdine, didanosine, docosanol, edoxudine, efavirenz,emtricitabine, enfuvirtide, entecavir, famciclovir, fomivirsen,fosamprenavir, foscarnet, fosfonet, fusion inhibitors (e.g.,enfuvirtide), ibacitabine, imunovir, idoxuridine, imiquimod, indinavir,inosine, integrase inhibitor, interferon type III, interferon type II,interferon type I, interferon, lamivudine, lopinavir, loviride,maraviroc, moroxydine, nelfinavir, nevirapine, nexavir, nucleosideanalogues, peginterferon alfa-2a, penciclovir, peramivir, pleconaril,podophyllotoxin, protease inhibitor, raltegravir, reverse transcriptaseinhibitor, ribavirin, rimantadine, ritonavir, pyrimidine antiviral,saquinavir, stavudine, synergistic enhancer (antiretroviral), tenofovir,tenofovir disoproxil, tipranavir, trifluridine, trizivir, tromantadine,truvada, valaciclovir (Valtrex™), vicriviroc, vidarabine, viramidine,zalcitabine, and zidovudine.

In a specific embodiment, the antiviral agent is acyclovir, ganciclovir,or valganciclovir.

In some embodiments, the antiviral agent is a nucleoside. Examples ofnucleoside analogs include acyclovir (ACV), ganciclovir (GCV),valganciclovir, famciclovir, foscarnet, ribavirin, zalcitabine (ddC),zidovudine (AZT), stavudine (D4T), larnivudine (3TC), didanosine (ddI),cytarabine, dideoxyadenosine, edoxudine, floxuridine, idozuridine,inosine pranobex, 2′-deoxy-5-(methylamino)uridine, trifluridine andvidarabine. Examples of a few protease inhibitors that show particularpromise in human therapy include saquinivir, ritonavir and indinavir.Other anti-viral agents include interferons (e.g. α-, β-, γ-interferon),cytokines such as tumor necrosis factor (TNF) or interleukins, cellreceptors and growth factor antagonists, which can be purified orrecombinantly produced.

In some embodiments, the antiviral agent is administered at a dose ofless than 3000 mg/day. In some embodiments, the antiviral agent isadministered at a dose of about 10 mg/day, about 20 mg/day, about 50mg/day, about 100 mg/day, about 150 mg/day, about 200 mg/day, about 250mg/day, about 300 mg/day, about 350 mg/day, about 400 mg/day, about 450mg/day, about 500 mg/day, about 600 mg/day, about 700 mg/day, about 800mg/day, about 900 mg/day, about 1000 mg/day, about 1200 mg/day, about1250 mg/day, about 1400 mg/day, about 1500 mg/day, about 1600 mg/day,about 1750 mg/day, about 1800 mg/day, about 1900 mg/day, about 2000mg/day, about 2250 mg/day, about 2500 mg/day, about 2750 mg/day, about3000 mg/day, about 3250 mg/day, about 350 0 mg/day, about 3750 mg/day,about 4000 mg/day, about 4250 mg/day, about 4500 mg/day, about 4750mg/day, or about 5000 mg/day. In certain embodiments, the antiviralagent is administered at a dose of less than 10 mg/day, less than 20mg/day, less than 50 mg/day, less than 100 mg/day, less than 150 mg/day,less than 200 mg/day, less than 250 mg/day, less than 300 mg/day, lessthan 350 mg/day, less than 400 mg/day, less than 450 mg/day, less than500 mg/day, less than 600 mg/day, less than 700 mg/day, less than 800mg/day, less than 900 mg/day, less than 1000 mg/day, less than 1200mg/day, less than 1250 mg/day, less than 1400 mg/day, less than 1500mg/day, less than 1600 mg/day, less than 1750 mg/day, less than 1800mg/day, less than 1900 mg/day, less than 2000 mg/day, less than 2250mg/day, less than 2500 mg/day, less than 2750 mg/day, less than 3000mg/day, less than 3250 mg/day, less than 3500 mg/day, less than 3750mg/day, less than 4000 mg/day, less than 4250 mg/day, less than 4500mg/day, less than 4750 mg/day, or less than 5000 mg/day. In someembodiments, the antiviral agent is administered at a dose of more than10 mg/day, more than 20 mg/day, more than 50 mg/day, more than 100mg/day, more than 150 mg/day, more than 200 mg/day, more than 250mg/day, more than 300 mg/day, more than 350 mg/day, more than 400mg/day, more than 450 mg/day, more than 500 mg/day, more than 600mg/day, more than 700 mg/day, more than 800 mg/day, more than 900mg/day, more than 1000 mg/day, more than 1200 mg/day, more than 1250mg/day, more than 1400 mg/day, more than 1500 mg/day, more than 1600mg/day, more than 1750 mg/day, more than 1800 mg/day, more than 1900mg/day, more than 2000 mg/day, more than 2250 mg/day, more than 2500mg/day, more than 2750 mg/day, more than 3000 mg/day, more than 3250mg/day, more than 3500 mg/day, more than 3750 mg/day, more than 4000mg/day, more than 4250 mg/day, more than 4500 mg/day, more than 4750mg/day, or more than 5000 mg/day. In certain embodiments, the antiviralagent is administered at a dose of more than 10 mg/day and less than5000 mg/day. In some embodiments, the antiviral agent is administered ata dose of more than 200 mg/day and less than 1000 mg/day. In certainembodiments, the antiviral agent is administered once a day (q.d.),twice a day (b.id.), or thrice a day (t.i.d.). In some embodiments, theantiviral agent is administered daily, once a week, twice a week, threetimes a week, four times a week, or five times a week.

In certain embodiments, the antiviral agent is ganciclovir. In someembodiments, ganciclovir is administered at a total daily dose of 3000mg/day. In certain embodiments, ganciclovir is administered at a dose of1000 mg three times a day. In some embodiments, ganciclovir isadministered at a dose of about 100 mg/day, about 250 mg/day, about 500mg/day, about 750 mg/day, about 1000 mg/day, about 1500 mg/day, about2000 mg/day, about 2500 mg/day, about 3000 mg/day, about 3500 mg/day, orabout 4000 mg/day. In certain embodiments, ganciclovir is administeredat a dose of less than 100 mg/day, less than 250 mg/day, less than 500mg/day, less than 750 mg/day, less than 1000 mg/day, less than 1500mg/day, less than 2000 mg/day, less than 2500 mg/day, less than 3000mg/day, less than 3500 mg/day, or less than 4000 mg/day. In someembodiments, ganciclovir is administered at a dose of more than 100mg/day, more than 250 mg/day, more than 500 mg/day, more than 750mg/day, more than 1000 mg/day, more than 1500 mg/day, more than 2000mg/day, more than 2500 mg/day, more than 3000 mg/day, more than 3500mg/day, or more than 4000 mg/day. In certain embodiments, ganciclovir isadministered at a dose of more than 500 mg/day and less 4000 mg/day. Insome embodiments, ganciclovir is administered at a dose of more than1000 mg/day and less than 3000 mg/day. In some embodiments, gancicloviris administered once a day, twice a day, or three times a day. Incertain embodiments, ganciclovir is administered once a week, twice aweek, three times a week, four times a week, five times a week, ordaily.

In some embodiments, the antiviral agent is valganciclovir. In certainembodiments, valganciclovir is administered at a total daily dose of 900mg/day. In some embodiments, valganciclovir is administered at a dose of900 mg once a day. In certain embodiments, valganciclovir isadministered at a total daily dose of 1800 mg/day. In some embodiments,valganciclovir is administered at a dose of 900 mg twice a day.

In some embodiments, valganciclovir is administered at a dose of about100 mg/day, about 200 mg/day, about 300 mg/day, about 400 mg/day, about500 mg/day, about 600 mg/day, about 700 mg/day, about 800 mg/day, about900 mg/day, about 1000 mg/day, about 1100 mg/day, about 1200 mg/day,about 1300 mg/day, about 1400 mg/day, about 1500 mg/day, about 1600mg/day, about 1700 mg/day, about 1800 mg/day, about 1900 mg/day, orabout 2000 mg/day. In certain embodiments, valganciclovir isadministered at a dose of less than 100 mg/day, less than 200 mg/day,less than 300 mg/day, less than 400 mg/day, less than 500 mg/day, lessthan 600 mg/day, less than 700 mg/day, less than 800 mg/day, less than900 mg/day, less than 1000 mg/day, less than 1100 mg/day, less than 1200mg/day, less than 1300 mg/day, less than 1400 mg/day, less than 1500mg/day, less than 1600 mg/day, less than 1700 mg/day, less than 1800mg/day, less than 1900 mg/day, or less than 2000 mg/day. In someembodiments, valganciclovir is administered at a dose of more than 100mg/day, more than 200 mg/day, more than 300 mg/day, more than 400mg/day, more than 500 mg/day, more than 600 mg/day, more than 700mg/day, more than 800 mg/day, more than 900 mg/day, more than 1000mg/day, more than 1100 mg/day, more than 1200 mg/day, more than 1300mg/day, more than 1400 mg/day, more than 1500 mg/day, more than 1600mg/day, more than 1700 mg/day, more than 1800 mg/day, more than 1900mg/day, or more than 2000 mg/day. In certain embodiments, valgancicloviris administered at a dose of more than 100 mg/day and less 2000 mg/day.In some embodiments, valganciclovir is administered at a dose of morethan 500 mg/day and less than 1500 mg/day. In some embodiments,valganciclovir is administered once a day, twice a day, or three times aday. In certain embodiments, valganciclovir is administered once a week,twice a week, three time a week, four times a week, five times a week,or daily.

In a specific embodiment, the antiviral agent is not a heat shockprotein inhibitor, an immunosuppressant, an antibiotic, aglucocorticoid, a non-steroidal anti-inflammatory drug, a Cox-2-specificinhibitor or a TNF-α binding protein. In a related embodiment, theantiviral agent is not a Hsp90 inhibitor, tacrolimus, cyclosporin,rapamycin (sirolimus), methotrexate, cyclophosphamide, azathioprine,mercaptopurine, mycophenolate, FTY720, levofloxacin, amoxycillin,prednisone, cortisone acetate, prednisolone, methylprednisolone,dexamethasone, betamethasone, triamcinolone, beclometasone,fludrocortisone acetate, deoxycorticosterone acetate, aldosterone,salicylates, arylalkanoic acids, a 2-arylpropionic acid, aN-arylanthranilic acid, an oxicam, a coxib, a sulphonanilide,valdecoxib, celecoxib, rofecoxib, leflunomide, gold thioglucose, goldthiomalate, aurofin, sulfasalazine, hydroxychloroquinine, minocycline,infliximab, etanercept, adalimumab, abatacept, anakinra, interferon-β,interferon-γ, interleukin-2, an allergy vaccine, an antihistamine, anantileukotriene, a beta-agonist, theophylline, or an anticholinergic.

Additional Agents

The methods of the provided invention can comprise administering to asubject a viral inducing agent, and antiviral agent, and one or moreadditional active agents. The additional agent can be selected based onthe type of viral, virally-induced, or inflammatory condition thesubject has or is suspected of having. The additional agent cancomprise, for example, another antiviral agent, another viral inducingagent, a vaccine, or an anticancer agent. For example, a subject withmultiple sclerosis can be administered a viral inducing agent, anantiviral agent, and a vaccine, for example, a vaccine comprising myelinbasic protein. In another example, a subject with diabetes can beadministered a viral inducing agent, an antiviral agent, and a vaccine,for example, a vaccine comprising an antigen.

In some embodiments, an additional agent is a anticancer agent. Incertain embodiments, the anticancer agent is a chemotherapeuticanticancer agent. Examples of chemotherapeutic anticancer agentsinclude, but are not limited to, nitrogen mustards; alkyl sulfonates;ethylene imines; nitrosoureas; epoxides; other alkylating agents; folicacid analogues; purine analogs; pyrimidine analogs; vinca alkaloids;podophyllotoxin derivatives; colchicine derivatives; taxanes; otherplant alkaloids and natural products; actinomycines; antracyclines;other cytotoxic antibiotics; platinum compounds; methylhydrazines;sensitizers; protein kinase inhibitors; other antineoplastic agents;estrogens; progestogens; gonadotropin releasing hormone analogs;anti-estrogens; anti-androgens; enzyme inhibitors; other hormoneantagonists; immunostimulants; immunosuppressants; calcineurininhibitors; and radiopharmaceuticals. In some embodiments, theanticancer agent is a toxin, e.g. diphtheria toxin.

In certain embodiments, an additional agent is a non-steroidalanti-inflammatory agent (NSAID). NSAID include, for example, Aspirin(Anacin™, Ascriptin™, Bayer™, Bufferin™, Ecotrin™, Excedrin™), Cholineand magnesium salicylates (CMT™, Tricosal™, Trilisate™), cholinesalicylate (Arthropan™), celecoxib (Celebrex™), diclofenac potassium(Cataflam™), diclofenac sodium (Voltaren™, Voltaren XR™), diclofenacsodium with misoprostol (Arthrotec™), diflunisal (Dolobid™), etodolac(Lodine™, Lodine XL™), fenoprofen calcium (Nalfon™), flurbiprofen(Ansaid™), ibuprofen (Advil™, Motrin™, Motrin IB™ Nuprin™), indomethacin(Indocin™, Indocin SR™), ketoprofen (Actron™, Orudis™, Orudis KT™,Oruvail™), magnesium salicylate (Arthritab™, Bayer Select™, Doan'sPills™, Magan™, Mobidin™ Mobogesic™), meclofenamate sodium (Meclomen™),mefenamic acid (Ponstel™), meloxicam (Mobic™), nabumetone (Relafen™),naproxen (Naprosyn™, Naprelan™), naproxen sodium (Aleve™, Anaprox™),oxaprozin (Daypro™), piroxicam (Feldene™), rofecoxib (Vioxx™), salsalate(Amigesic™, Anaflex 750™, Disalcid™, Marthritic™, Mono-Gesic™, Salflex™,Salsitab™), sodium salicylate (various generics), sulindac (Clinoril™),tolmetin sodium (Tolectin™), valdecoxib (Bextra™).

In some embodiments, an additional agent is a lipid lowering agent. Incertain embodiments, the lipid lowering agent is a statin. Examples ofstatins include, but are not limited to, Advicor® (niacinextended-release/lovastatin), Altoprev® (lovastatin extended-release),Caduet® (amlodipine and atorvastatin), Crestor® (rosuvastatin), Lescol®(fluvastatin), Lescol XL (fluvastatin extended-release), Lipitor®(atorvastatin), Mevacor® (lovastatin), Pravachol® (pravastatin), Simcor®(niacin extended-release/simvastatin), Vytorin® (ezetimibe/simvastatin),and Zocor® (simvastatin). A lipid lowering agent can be administered toa subject that has or is suspected of having atherosclerosis. Forexample, a subject with cytomegalovirus induced atherosclerosis can beadministered an additional agent that can comprise atorvastatin,rosuvastatin, lovastatin, simvastatin, or pravastatin.

In certain embodiments, an additional agent is an immunosuppressivedrug. Immunosuppressive drug, for example, include glucocorticoids,antibodies, cytostatic agents, and drugs that act on immunophilins.Glucocorticoids can include, for example, prednisolone, prednisone, ormethylprednisolone. A cytostatic agent can include, for example, anagent that interferes with nucleic acid synthesis, for example, folicacid, pyrimidine analogs, and purine analogs. A folic acid analog thatcan be used as an immunosuppressive drug is methotrexate, which can binddihydrofolate reductase and prevent the synthesis of tetrahydrofolate.Another cytostatic agent is azathioprine, which can be cleavednonezymatically to form mercaptopurine, which can act as a purineanalogue. A cytostatic agent can include, for example, an alkylatingagent, including, for example, cyclophosphamide, and a nitrosourea. Acytostatic agent can be a platinum compound. Other cytostatic agentsinclude, for example, cytotoxic antibiotics, including dactinomycin,anthracylcines, mitomycin C, bleomycin, and mithramycin. Examples ofantibodies that can be immunosuppressive agents include, for example,heterologous polyclonal antibodies, for example, from rabbit or horse.Other antibodies include monoclonal antibodies directed to specificantigens e.g., T-cell receptor directed antibodies (e.g., OKT3,muromonab, which targets CD3), and IL-2 receptor directed antibodies(e.g., targeting CD25). Drugs that can act on immunophilins include, forexample, cyclosporin, tacrolimus (Prograf), Sirolimus (rapamycin,Rapamune). Other drugs that can act as immunosuppressive drugs include,for example, mycophenolate (mycophenolic acid), interferons, opioids,TNF binding proteins, Fingolimod, myriocin, and ciclosporin.

The additional agent can be, for example, FK506, a monoclonal antibody,an anti-T cell monoclonal antibody, an anti-B cell monoclonal antibody,or a TNF inhibitor. The monoclonal antibody can be an anti-B cellantibody. The anti-B cell antibody can be anti-CD20. The TNF inhibitorcan be infliximab (Remicade™), etanercept (Enbrel™), adalimumab(Humira™), or an anti-IL-6 antibody.

A subject with an autoimmune condition can be administered a viralinducing agent, an antiviral agent, and an additional agent, where theadditional agent comprises cyclosporine, azathiorprine, methotrexate,cyclophosphamide, FK506, tacrolimus, a monoclonal antibody, an anti-Tcell monoclonal antibody, an anti-B cell monoclonal antibody, an IL-2receptor antibody, or a TNF inhibitor.

The additional agent can be glatiramer (Copaxone™), Natalizumab(Tysabri™), mitoxantrone (Novantrone™), cladribine, or Campath antibody.For example, a subject with multiple sclerosis can be administered anadditional agent that can comprise glatiramer, mitoxantrone,natalizumab, cladribine, or Campath antibody.

Types of Viruses and Virally-Induced Conditions

The methods and compositions provided herein can be used to treat and/orprevent viral infections. The virus causing the infection can be amember of the herpes virus family, a human immunodeficiency virus,parvovirus, or coxsackie virus. A member of the herpes virus family canbe herpes simplex virus, herpes genitalis virus, varicella zoster virus,Epstein-Barr virus, human herpes virus 6, human herpes virus 8, orcytomegalovirus. The subject can have coronary artery conditionassociated with a cytomegalovirus or herpes simplex virus infection. Thesubject can have autoimmune condition associated with Epstein-Barr virusinfection. The subject can have a lymphoma or other cancer associatedwith Epstein-Barr virus infection.

In some embodiments, the viral or virally-induced condition is caused bya retrovirus, such as HIV, HTLV1 and 2. In certain embodiments, theviral or virally-induced condition is caused by a DNA virus, such as aherpesvirus. In some embodiments, the herpesvirus is an Epstein-Barrvirus, cytomegalovirus, Herpes type 1 (oral herpes), herpes type 2,Kaposi's sarcoma virus (human herpes virus 8), BK viruses, or hepatitisvirus. In certain embodiments, the virally-induced or virus-associateddisease is a cancer. In some embodiments, the virally-induced orvirus-associated cancer is a lymphoma, chronic lyphocytic leukemia,nasopharyngeal carcinoma, gastric cancer, or Kaposi's sarcoma. In otherembodiments, the virally-induced or virus-associated disease is anautoimmune disease. In certain embodiments, the autoimmune disease isrheumatoid arthritis, systemic lupus erythematosus, or multiplesclerosis.

The methods and compositions described herein can be used to treatand/or prevent infections caused by any virus, including, for example,Abelson leukemia virus, Abelson murine leukemia virus, Abelson's virus,Acute laryngotracheobronchitis virus, Adelaide River virus, Adenoassociated virus group, Adenovirus, African horse sickness virus,African swine fever virus, AIDS virus, Aleutian mink conditionparvovirus, Alpharetrovirus, Alphavirus, ALV related virus, Amaparivirus, Aphthovirus, Aquareovirus, Arbovirus, Arbovirus C, arbovirusgroup A, arbovirus group B, Arenavirus group, Argentine hemorrhagicfever virus, Argentine hemorrhagic fever virus, Arterivirus, Astrovirus,Ateline herpesvirus group, Aujezky's condition virus, Aura virus, Ausdukcondition virus, Australian bat lyssavirus, Aviadenovirus, avianerythroblastosis virus, avian infectious bronchitis virus, avianleukemia virus, avian leukosis virus, avian lymphomatosis virus, avianmyeloblastosis virus, avian paramyxovirus, avian pneumoencephalitisvirus, avian reticuloendotheliosis virus, avian sarcoma virus, aviantype C retrovirus group, Avihepadnavirus, Avipoxvirus, B virus, B19virus, Babanki virus, baboon herpesvirus, baculovirus, Barmah Forestvirus, Bebaru virus, Berrimah virus, Betaretrovirus, Birnavirus, Bittnervirus, BK virus, Black Creek Canal virus, bluetongue virus, Bolivianhemorrhagic fever virus, Boma condition virus, border condition of sheepvirus, borna virus, bovine alphaherpesvirus 1, bovine alphaherpesvirus2, bovine coronavirus, bovine ephemeral fever virus, bovineimmunodeficiency virus, bovine leukemia virus, bovine leukosis virus,bovine mammillitis virus, bovine papillomavirus, bovine papularstomatitis virus, bovine parvovirus, bovine syncytial virus, bovine typeC oncovirus, bovine viral diarrhea virus, Buggy Creek virus, bulletshaped virus group, Bunyamwera virus supergroup, Bunyavirus, Burkitt'slymphoma virus, Bwamba Fever, CA virus, Calicivirus, Californiaencephalitis virus, camelpox virus, canarypox virus, canid herpesvirus,canine coronavirus, canine distemper virus, canine herpesvirus, canineminute virus, canine parvovirus, Cano Delgadito virus, caprine arthritisvirus, caprine encephalitis virus, Caprine Herpes Virus, Capripox virus,Cardiovirus, caviid herpesvirus 1, Cercopithecid herpesvirus 1,cercopithecine herpesvirus 1, Cercopithecine herpesvirus 2, Chandipuravirus, Changuinola virus, channel catfish virus, Charleville virus,chickenpox virus, Chikungunya virus, chimpanzee herpesvirus, chubreovirus, chum salmon virus, Cocal virus, Coho salmon reovirus, coitalexanthema virus, Colorado tick fever virus, Coltivirus, Columbia SKvirus, common cold virus, contagious ecthyma virus, contagious pustulardermatitis virus, Coronavirus, Corriparta virus, coryza virus, cowpoxvirus, coxsackie virus, CPV (cytoplasmic polyhedrosis virus), cricketparalysis virus, Crimean-Congo hemorrhagic fever virus, croup associatedvirus, Cryptovirus, Cypovirus, Cytomegalovirus, cytomegalovirus group,cytoplasmic polyhedrosis virus, deer papillomavirus, deltaretrovirus,dengue virus, Densovirus, Dependovirus, Dhori virus, diploma virus,Drosophila C virus, duck hepatitis B virus, duck hepatitis virus 1, duckhepatitis virus 2, duovirus, Duvenhage virus, Deformed wing virus DWV,eastern equine encephalitis virus, eastern equine encephalomyelitisvirus, EB virus, Ebola virus, Ebola-like virus, echo virus, echovirus,echovirus 10, echovirus 28, echovirus 9, ectromelia virus, EEE virus,EIA virus, EIA virus, encephalitis virus, encephalomyocarditis groupvirus, encephalomyocarditis virus, Enterovirus, enzyme elevating virus,enzyme elevating virus (LDH), epidemic hemorrhagic fever virus,epizootic hemorrhagic condition virus, Epstein-Barr virus, equidalphaherpesvirus 1, equid alphaherpesvirus 4, equid herpesvirus 2,equine abortion virus, equine arteritis virus, equine encephalosisvirus, equine infectious anemia virus, equine morbillivirus, equinerhinopneumonitis virus, equine rhinovirus, Eubenangu virus, European elkpapillomavirus, European swine fever virus, Everglades virus, Eyachvirus, felid herpesvirus 1, feline calicivirus, feline fibrosarcomavirus, feline herpesvirus, feline immunodeficiency virus, felineinfectious peritonitis virus, feline leukemia/sarcoma virus, felineleukemia virus, feline panleukopenia virus, feline parvovirus, felinesarcoma virus, feline syncytial virus, Filovirus, Flanders virus,Flavivirus, foot and mouth condition virus, Fort Morgan virus, FourCorners hantavirus, fowl adenovirus 1, fowlpox virus, Friend virus,Gammaretrovirus, GB hepatitis virus, GB virus, German measles virus,Getah virus, gibbon ape leukemia virus, glandular fever virus, goatpoxvirus, golden shinner virus, Gonometa virus, goose parvovirus,granulosis virus, Gross' virus, ground squirrel hepatitis B virus, groupA arbovirus, Guanarito virus, guinea pig cytomegalovirus, guinea pigtype C virus, Hantaan virus, Hantavirus, hard clam reovirus, harefibroma virus, HCMV (human cytomegalovirus), hemadsorption virus 2,hemagglutinating virus of Japan, hemorrhagic fever virus, hendra virus,Henipaviruses, Hepadnavirus, hepatitis A virus, hepatitis B virus group,hepatitis C virus, hepatitis D virus, hepatitis delta virus, hepatitis Evirus, hepatitis F virus, hepatitis G virus, hepatitis nonA nonB virus,hepatitis virus, hepatitis virus (nonhuman), hepatoencephalomyelitisreovirus 3, Hepatovirus, heron hepatitis B virus, herpes B virus, herpessimplex virus, herpes simplex virus 1, herpes simplex virus 2,herpesvirus, herpesvirus 7, Herpesvirus ateles, Herpesvirus hominis,Herpesvirus infection, Herpesvirus saimiri, Herpesvirus suis,Herpesvirus varicellae, Highlands J virus, Hirame rhabdovirus, hogcholera virus, human adenovirus 2, human alphaherpesvirus 1, humanalphaherpesvirus 2, human alphaherpesvirus 3, human B lymphotropicvirus, human betaherpesvirus 5, human coronavirus, human cytomegalovirusgroup, human foamy virus, human gammaherpesvirus 4, humangammaherpesvirus 6, human hepatitis A virus, human herpesvirus 1 group,human herpesvirus 2 group, human herpesvirus 3 group, human herpesvirus4 group, human herpesvirus 6, human herpesvirus 8, humanimmunodeficiency virus, human immunodeficiency virus 1, humanimmunodeficiency virus 2, human papillomavirus, human T cell leukemiavirus, human T cell leukemia virus I, human T cell leukemia virus II,human T cell leukemia virus III, human T cell lymphoma virus I, human Tcell lymphoma virus II, human T cell lymphotropic virus type 1, human Tcell lymphotropic virus type 2, human T lymphotropic virus I, human Tlymphotropic virus II, human T lymphotropic virus III, Ichnovirus,infantile gastroenteritis virus, infectious bovine rhinotracheitisvirus, infectious haematopoietic necrosis virus, infectious pancreaticnecrosis virus, influenza virus A, influenza virus B, influenza virus C,influenza virus D, influenza virus pr8, insect iridescent virus, insectvirus, iridovirus, Japanese B virus, Japanese encephalitis virus, JCvirus, Junin virus, Kaposi's sarcoma-associated herpesvirus, Kemerovovirus, Kilham's rat virus, Klamath virus, Kolongo virus, Koreanhemorrhagic fever virus, kumba virus, Kysanur forest condition virus,Kyzylagach virus, La Crosse virus, lactic dehydrogenase elevating virus,lactic dehydrogenase virus, Lagos bat virus, Langur virus, lapineparvovirus, Lassa fever virus, Lassa virus, latent rat virus, LCM virus,Leaky virus, Lentivirus, Leporipoxvirus, leukemia virus, leukovirus,lumpy skin condition virus, lymphadenopathy associated virus,Lymphocryptovirus, lymphocytic choriomeningitis virus,lymphoproliferative virus group, Machupo virus, mad itch virus,mammalian type B oncovirus group, mammalian type B retroviruses,mammalian type C retrovirus group, mammalian type D retroviruses,mammary tumor virus, Mapuera virus, Marburg virus, Marburg-like virus,Mason Pfizer monkey virus, Mastadenovirus, Mayaro virus, ME virus,measles virus, Menangle virus, Mengo virus, Mengovirus, Middelburgvirus, milkers nodule virus, mink enteritis virus, minute virus of mice,MLV related virus, MM virus, Mokola virus, Molluscipoxvirus, Molluscumcontagiosum virus, monkey B virus, monkeypox virus, Mononegavirales,Morbillivirus, Mount Elgon bat virus, mouse cytomegalovirus, mouseencephalomyelitis virus, mouse hepatitis virus, mouse K virus, mouseleukemia virus, mouse mammary tumor virus, mouse minute virus, mousepneumonia virus, mouse poliomyelitis virus, mouse polyomavirus, mousesarcoma virus, mousepox virus, Mozambique virus, Mucambo virus, mucosalcondition virus, mumps virus, murid betaherpesvirus 1, muridcytomegalovirus 2, murine cytomegalovirus group, murineencephalomyelitis virus, murine hepatitis virus, murine leukemia virus,murine nodule inducing virus, murine polyomavirus, murine sarcoma virus,Muromegalovirus, Murray Valley encephalitis virus, myxoma virus,Myxovirus, Myxovirus multiforme, Myxovirus parotitidis, Nairobi sheepcondition virus, Nairovirus, Nanirnavirus, Nariva virus, Ndumo virus,Neethling virus, Nelson Bay virus, neurotropic virus, New WorldArenavirus, newborn pneumonitis virus, Newcastle condition virus, Nipahvirus, noncytopathogenic virus, Norwalk virus, nuclear polyhedrosisvirus (NPV), nipple neck virus, O'nyong'nyong virus, Ockelbo virus,oncogenic virus, oncogenic viruslike particle, oncornavirus, Orbivirus,Orf virus, Oropouche virus, Orthohepadnavirus, Orthomyxovirus,Orthopoxvirus, Orthoreovirus, Orungo, ovine papillomavirus, ovinecatarrhal fever virus, owl monkey herpesvirus, Palyam virus,Papillomavirus, Papillomavirus sylvilagi, Papovavirus, parainfluenzavirus, parainfluenza virus type 1, parainfluenza virus type 2,parainfluenza virus type 3, parainfluenza virus type 4, Paramyxovirus,Parapoxvirus, paravaccinia virus, Parvovirus, Parvovirus B19, parvovirusgroup, Pestivirus, Phlebovirus, phocine distemper virus, Picodnavirus,Picornavirus, pig cytomegalovirus—pigeonpox virus, Piry virus, Pixunavirus, pneumonia virus of mice, Pneumovirus, poliomyelitis virus,poliovirus, Polydnavirus, polyhedral virus, polyoma virus, Polyomavirus,Polyomavirus bovis, Polyomavirus cercopitheci, Polyomavirus hominis 2,Polyomavirus maccacae 1, Polyomavirus muris 1, Polyomavirus muris 2,Polyomavirus papionis 1, Polyomavirus papionis 2, Polyomavirussylvilagi, Pongine herpesvirus 1, porcine epidemic diarrhea virus,porcine hemagglutinating encephalomyelitis virus, porcine parvovirus,porcine transmissible gastroenteritis virus, porcine type C virus, poxvirus, poxvirus, poxvirus variolae, Prospect Hill virus, Provirus,pseudocowpox virus, pseudorabies virus, psittacinepox virus, quailpoxvirus, rabbit fibroma virus, rabbit kidney vaculolating virus, rabbitpapillomavirus, rabies virus, raccoon parvovirus, raccoonpox virus,Ranikhet virus, rat cytomegalovirus, rat parvovirus, rat virus,Rauscher's virus, recombinant vaccinia virus, recombinant virus,reovirus, reovirus 1, reovirus 2, reovirus 3, reptilian type C virus,respiratory infection virus, respiratory syncytial virus, respiratoryvirus, reticuloendotheliosis virus, Rhabdovirus, Rhabdovirus carpia,Rhadinovirus, Rhinovirus, Rhizidiovirus, Rift Valley fever virus,Riley's virus, rinderpest virus, RNA tumor virus, Ross River virus,Rotavirus, rougeole virus, Rous sarcoma virus, rubella virus, rubeolavirus, Rubivirus, Russian autumn encephalitis virus, SA 11 simian virus,SA2 virus, Sabia virus, Sagiyama virus, Saimirine herpesvirus 1,salivary gland virus, sandfly fever virus group, Sandjimba virus, SARSvirus, SDAV (sialodacryoadenitis virus), sealpox virus, Semliki ForestVirus, Seoul virus, sheeppox virus, Shope fibroma virus, Shope papillomavirus, simian foamy virus, simian hepatitis A virus, simian humanimmunodeficiency virus, simian immunodeficiency virus, simianparainfluenza virus, simian T cell lymphotrophic virus, simian virus,simian virus 40, Simplexvirus, Sin Nombre virus, Sindbis virus, smallpoxvirus, South American hemorrhagic fever viruses, sparrowpox virus,Spumavirus, squirrel fibroma virus, squirrel monkey retrovirus, SSV 1virus group, STLV (simian T lymphotropic virus) type I, STLV (simian Tlymphotropic virus) type II, STLV (simian T lymphotropic virus) typeIII, stomatitis papulosa virus, submaxillary virus, suidalphaherpesvirus 1, suid herpesvirus 2, Suipoxvirus, swamp fever virus,swinepox virus, Swiss mouse leukemia virus, TAC virus, Tacaribe complexvirus, Tacaribe virus, Tanapox virus, Taterapox virus, Tench reovirus,Theiler's encephalomyelitis virus, Theiler's virus, Thogoto virus,Thottapalayam virus, Tick borne encephalitis virus, Tioman virus,Togavirus, Torovirus, tumor virus, Tupaia virus, turkey rhinotracheitisvirus, turkeypox virus, type C retroviruses, type D oncovirus, type Dretrovirus group, ulcerative condition rhabdovirus, Una virus, Uukuniemivirus group, vaccinia virus, vacuolating virus, varicella zoster virus,Varicellovirus, Varicola virus, variola major virus, variola virus,Vasin Gishu condition virus, VEE virus, Venezuelan equine encephalitisvirus, Venezuelan equine encephalomyelitis virus, Venezuelan hemorrhagicfever virus, vesicular stomatitis virus, Vesiculovirus, Vilyuisk virus,viper retrovirus, viral haemorrhagic septicemia virus, Visna Maedivirus, Visna virus, volepox virus, VSV (vesicular stomatitis virus),Wallal virus, Warrego virus, wart virus, WEE virus, West Nile virus,western equine encephalitis virus, western equine encephalomyelitisvirus, Whataroa virus, Winter Vomiting Virus, woodchuck hepatitis Bvirus, woolly monkey sarcoma virus, wound tumor virus, WRSV virus, Yabamonkey tumor virus, Yaba virus, Yatapoxvirus, yellow fever virus, andthe Yug Bogdanovac virus.

Inflammatory Conditions

Inflammatory conditions that can be treated and/or prevented using themethods and compositions provided herein include, for example,autoimmune condition. Autoimmune conditions include, for example,rheumatoid arthritis, multiple sclerosis, Sjogren's syndrome, systemiclupus erythematosus, autoimmune hepatitis, autoimmune thyroiditis,hemophagocytic syndrome (hemophagocytic lymphohistiocytosis), diabetesmellitus type 1, Crohn's condition, ulcerative colitis, psoriasis,psoriatic arthritis, idiopathic thrombocytonpenic pupura, polymyositis,dermatomyositis, myasthenia gravis, autoimmune thryroiditis, Evan'ssyndrome, autoimmune hemolytic anemia, aplastic anemia, autoimmuneneutropenia, scleroderma, Reiter's syndrome, ankylosing spondylitis,pemphnigus, pemphigoid or autoimmune hepatitis, Behçet's condition,Celiac condition, Chagas condition, acute disseminatedencephalomyelitis, Addison's condition, antiphospholipid antibodysyndrome, autoimmune inner ear condition, bullous pemphigoid, Chronicobstructive pulmonary condition, Goodpasture's syndrome, Graves'condition, Guillain-Barré syndrome, Hashimoto's thyroditis, Hidradenitissuppurativa, Interstitial cystitis, neuromyotonia, pemphigus vulgaris,pernicious anemia, primary biliary cirrhosis, and vasculitis syndromes.

Other inflammatory conditions that can be treated and/or prevented usingthe methods and compositions of the provided invention include, forexample, an allergic condition (e.g., allergic rhinitis, asthma, atopiceczema), a skin condition, coronary artery condition, peripheral arterycondition, atherosclerosis, retinitis, pancreatitis, cardiomyopathy,pericarditis, colitis, glomerulonephritis, lung inflammation,esophagitis, gastritis, duodenitis, ileitis, meningitis, encephalitis,encephalomyelitis, transverse myelitis, cystitis, urethritis, mucositis,lymphadenitis, dermatitis, hepatitis, osteomyelitis, or herpes zoster.

Formulations, Routes of Administration, and Effective Doses

Another aspect of the present invention relates to formulations, routesof administration and effective doses for pharmaceutical compositionscomprising an agent or combination of agents. Such pharmaceuticalcompositions can be used to treat a virus-induced inflammatory conditionas described above. A pharmaceutical composition can comprise a viralinducing agent. A pharmaceutical composition can comprise a viralinducing agent and one or more additional agents. A pharmaceuticalcomposition can comprise an antiviral agent. A pharmaceuticalcomposition can comprise an antiviral agent and one or more additionalagents. A pharmaceutical composition can comprise a viral inducing agentand an antiviral agent. A pharmaceutical composition can comprise aviral inducing agent, an antiviral agent, and one or more additionalagents.

The agents or their pharmaceutically acceptable salts can be providedalone or in combination with one or more other agents or with one ormore other forms. For example, a formulation can comprise one or moreagents in particular proportions, depending on the relative potencies ofeach agent and the intended indication. For example, in compositions fortargeting two different targets and where potencies are similar, about a1:1 ratio of agents can be used. The two forms can be formulatedtogether, in the same dosage unit e.g. in one cream, suppository,tablet, capsule, enteric coated tablet or capsule, aerosol spray, orpacket of powder to be dissolved in a beverage; or each form may beformulated in a separate unit, e.g., two creams, two suppositories, twotablets, two capsules, a tablet and a liquid for dissolving the tablet,two aerosol sprays, or a packet of powder and a liquid for dissolvingthe powder, etc.

A “pharmaceutically acceptable salt” can be a salt that retains thebiological effectiveness and properties of one or more agents, and whichare not biologically or otherwise undesirable. For example, apharmaceutically acceptable salt does not interfere with the beneficialeffect of a viral inducing agent or an antiviral agent.

Salts can include those of the inorganic ions, for example, sodium,potassium, calcium, magnesium ions, and the like. Salts can includesalts with inorganic or organic acids, for example, hydrochloric acid,hydrobromic acid, phosphoric acid, nitric acid, sulfuric acid,methanesulfonic acid, p-toluenesulfonic acid, acetic acid, fumaric acid,succinic acid, lactic acid, mandelic acid, malic acid, citric acid,tartaric acid or maleic acid. If one or more agents contain a carboxygroup or other acidic group, it can be converted into a pharmaceuticallyacceptable addition salt with inorganic or organic bases. Examples ofsuitable bases include sodium hydroxide, potassium hydroxide, ammonia,cyclohexylamine, dicyclohexyl-amine, ethanolamine, diethanolamine,triethanolamine, and the like.

A pharmaceutically acceptable ester or amide can be an ester or amidethat retains biological effectiveness and properties of one or moreagents, and which are not biologically or otherwise undesirable. Forexample, the ester or amide does not interfere with the beneficialeffect of a viral inducing agent, an antiviral agent, or an additionalagent. Esters can include, for example, ethyl, methyl, isobutyl,ethylene glycol, and the like. Amides include can include, for example,unsubstituted amides, alkyl amides, dialkyl amides, and the like.

A viral inducing agent, for example a HDAC inhibitor, can beadministered in combination with an antiviral agent. Pharmaceuticalcompositions comprising a combination of a viral inducing agent and anantiviral agent can be formulated to comprise certain molar ratios. Forexample, molar ratios of about 99:1 to about 1:99 of a viral inducingagent to the antiviral agent can be used. The range of molar ratios ofviral inducing agent:the antiviral agent can be selected from about80:20 to about 20:80; about 75:25 to about 25:75, about 70:30 to about30:70, about 66:33 to about 33:66, about 60:40 to about 40:60; about50:50; and about 90:10 to about 10:90. The viral inducing agent and theantiviral agent can be co-formulated, in the same dosage unit, e.g., inone cream, suppository, tablet, capsule, enteric coated capsule ortablet, or packet of powder to be dissolved in a beverage; or eachagent, form, and/or compound can be formulated in separate units, e.g.,two creams, suppositories, tablets, two capsules, enteric coatedcapsules or tablets, a tablet and a liquid for dissolving the tablet, anaerosol spray a packet of powder and a liquid for dissolving the powder,etc.

An agent can be administered in combination with one or more othercompounds, forms, and/or agents, e.g., as described above.Pharmaceutical compositions comprising combinations of a viral inducingagent and/or antiviral agent with one or more other active agents can beformulated to comprise certain molar ratios. For example, molar ratiosof about 99:1 to about 1:99 of a viral inducing agent to the otheractive agent can be used; molar ratios of about 99:1 to about 1:99 of anantiviral agent to the other active agent can be used; molar ratios ofabout 99:1 to about 1:99 of a viral inducing agent and antiviral agentcan be used. The range of molar ratios of viral inducing agent:otheractive agent can be selected from about 80:20 to about 20:80; about75:25 to about 25:75, about 70:30 to about 30:70, about 66:33 to about33:66, about 60:40 to about 40:60; about 50:50; and about 90:10 to about10:90. The range of molar ratios of an antiviral agent:other activeagent can be selected from about 80:20 to about 20:80; about 75:25 toabout 25:75, about 70:30 to about 30:70, about 66:33 to about 33:66,about 60:40 to about 40:60; about 50:50; and about 90:10 to about 10:90.The molar ratio may of a viral inducing agent:other active agent can beabout 1:9 or about 1:1. The molar ratio may of an antiviral agent:otheractive agent can be about 1:9 or about 1:1. Two or more agents, formsand/or compounds can be formulated together, in the same dosage unit,e.g., in one cream, suppository, tablet, capsule, enteric coated capsuleor tablet, or packet of powder to be dissolved in a beverage; or eachagent, form, and/or compound can be formulated in separate units, e.g.,two creams, suppositories, tablets, two capsules, enteric coatedcapsules or tablets, a tablet and a liquid for dissolving the tablet, anaerosol spray a packet of powder and a liquid for dissolving the powder,etc.

A viral inducing agent, for example a HDAC inhibitor, can beadministered in combination with an antiviral agent. Pharmaceuticalcompositions comprising a combination of a viral inducing agent and anantiviral agent can be formulated to comprise certain mg per dose. Forexample, a viral inducing agent can be administered at 0.01, 0.05, 0.1,0.5, 1, 2, 5, 10, 20, 25, 50, 100, 250, 500, 1000 mg/kg per dose. A HDACinhibitor can be administered at 0.01-0.1, 0.05-0.5, 1-2, 1-5, 5-10,10-20, 10-25, 10-50, 100-500, or 500-1000 mg/kg per dose. A single doseof an oral formulation of a viral inducing agent can contain 0.01, 0.05,0.1, 0.5, 1, 2, 5, 10, 20, 25, 50, 100, 250, 500, 1000 mg. In oneembodiment, the HDAC inhibitor is administered at 0.01, 0.05, 0.1, 0.5,1, 2, 5, 10, 20, 25, 50, 100, 250, 500, 1000 mg/kg per dose. In arelated embodiment, the HDAC inhibitor is administered orally. In aspecific embodiment, the total daily oral dose of a HDAC inhibitor is nomore than 1, 2, 5, 10, 20, 25, 40, 50, 100, 250, or 500 mg. In anotherrelated embodiment, the HDAC inhibitor is administered 1, 2, 3, 4, or 5times a day orally. In other embodiments, a single daily dose of a HDACinhibitor is provided whereas oral valganciclovir is provided at 900mgs/dose, two times a day.

An oral formulation of an HDAC inhibitor can be co-formulated with anantiviral agent, such as valganciclovir. In a specific embodiment whenan HDAC inhibitor and valganciclovir are co-formulated for a singledaily dose, the valganciclovir is present in a slow release or timedrelease form. In certain embodiments, the HDAC inhibitor andvalganciclovir or other antiviral agent are co-formulated such that theHDAC inhibitor is present at no more than 100 mg per dose, and theantiviral agent is present at no more than 1000 mg per dose. In someembodiments, the HDAC inhibitor and valganciclovir or other antiviralagent are co-formulated such that the HDAC inhibitor is present at nomore than 80 mg per dose, and the antiviral agent is present at no morethan 500 mg per dose. In certain embodiments, the HDAC inhibitor andvalganciclovir or other antiviral agent are co-formulated such that theHDAC inhibitor is present at not greater than 80 mg per dose, and theantiviral agent is present at not greater than 1500 mg per dose.

In some embodiments, a co-formulation comprising an HDAC inhibitor andan antiviral agent comprises less than 500 mg, less than 400 mg, lessthan 300 mg, less than 200 mg, less than 100 mg, less than 90 mg, lessthan 80 mg, less than 70 mg, less than 60 mg, less than 50 mg, less than40 mg, less than 30 mg, less than 20 mg, less than 10 mg, less than 5mg, less than 2 mg, or less than 1 mg of the HDAC inhibitor. In otherembodiments, a co-formulation comprising an HDAC inhibitor and anantiviral agent comprises less than 1500 mg, less than 1400 mg, lessthan 1300 mg, less than 1200 mg, less than 1100 mg, less than 1000 mg,less than 900 mg, less than 800 mg, less than 700 mg, less than 600 mg,less than 500 mg, less than 400 mg, less than 300 mg, less than 200 mg,less than 100 mg, less than 90 mg, less than 80 mg, less than 70 mg,less than 60 mg, less than 50 mg, less than 40 mg, less than 30 mg, lessthan 20 mg, less than 10 mg, less than 5 mg, less than 2 mg, or lessthan 1 mg of the antiviral agent.

In certain embodiments, a unit dose of a co-formulated HDAC inhibitorand antiviral agent comprises between about 1 mg and about 500 mg of theHDAC inhibitor and between 1 mg and 1500 mg of the antiviral agent. Insome embodiments, the unit dose comprises about 500 mg, about 400 mg,about 300 mg, about 200 mg, about 100 mg, about 90 mg, about 80 mg,about 70 mg, about 60 mg, about 50 mg, about 40 mg, about 30 mg, about20 mg, about 10 mg, about 5 mg, about 2 mg, or about 1 mg of the HDACinhibitor. In certain embodiments, the unit dose comprises less than 500mg, less than 400 mg, less than 300 mg, less than 200 mg, less than 100mg, less than 90 mg, less than 80 mg, less than 70 mg, less than 60 mg,less than 50 mg, less than 40 mg, less than 30 mg, less than 20 mg, lessthan 10 mg, less than 5 mg, less than 2 mg, or less than 1 mg of an HDACinhibitor. In some embodiments, the unit dose comprises more than 500mg, more than 400 mg, more than 300 mg, more than 200 mg, more than 100mg, more than 90 mg, more than 80 mg, more than 70 mg, more than 60 mg,more than 50 mg, more than 40 mg, more than 30 mg, more than 20 mg, morethan 10 mg, more than 5 mg, more than 2 mg, or more than 1 mg of an HDACinhibitor. In certain embodiments, the unit dose comprises more than 2mg and less than 500 mg of an HDAC inhibitor. In some embodiments, theunit dose comprises more than 10 mg and less than 50 mg of an HDACinhibitor.

In some embodiments, the unit dose comprises about 2000 mg, about 1900mg, about 1800 mg, about 1700 mg, about 1600 mg, comprises about 1500mg, about 1400 mg, about 1300 mg, about 1200 mg, about 1100 mg, about1000 mg, about 900 mg, about 800 mg, about 750 mg, about 700 mg, about650 mg, about 600 mg, about 550 mg, about 500 mg, about 450 mg, about400 mg, about 350 mg, about 300 mg, about 250 mg, about 200 mg, about150 mg, about 140 mg, about 130 mg, about 120 mg, about 110 mg, about100 mg, about 90 mg, about 80 mg, about 70 mg, about 60 mg, about 50 mg,about 40 mg, about 30 mg, about 20 mg, about 10 mg, about 5 mg, about 2mg, or about 1 mg of the antiviral agent. In certain embodiments, theunit dose comprises less than 2000 mg, less than 1900 mg, less than 1800mg, less than 1700 mg, less than 1600 mg, comprises less than 1500 mg,less than 1400 mg, less than 1300 mg, less than 1200 mg, less than 1100mg, less than 1000 mg, less than 900 mg, less than 800 mg, less than 750mg, less than 700 mg, less than 650 mg, less than 600 mg, less than 550mg, less than 500 mg, less than 450 mg, less than 400 mg, less than 350mg, less than 300 mg, less than 250 mg, less than 200 mg, less than 150mg, less than 140 mg, less than 130 mg, less than 120 mg, less than 110mg, less than 100 mg, less than 90 mg, less than 80 mg, less than 70 mg,less than 60 mg, less than 50 mg, less than 40 mg, less than 30 mg, lessthan 20 mg, less than 10 mg, less than 5 mg, less than 2 mg, or lessthan 1 mg of the antiviral agent. In some embodiments, the unit dosecomprises more than 2000 mg, more than 1900 mg, more than 1800 mg, morethan 1700 mg, more than 1600 mg, comprises more than 1500 mg, more than1400 mg, more than 1300 mg, more than 1200 mg, more than 1100 mg, morethan 1000 mg, more than 900 mg, more than 800 mg, more than 750 mg, morethan 700 mg, more than 650 mg, more than 600 mg, more than 550 mg, morethan 500 mg, more than 450 mg, more than 400 mg, more than 350 mg, morethan 300 mg, more than 250 mg, more than 200 mg, more than 150 mg, morethan 140 mg, more than 130 mg, more than 120 mg, more than 110 mg, morethan 100 mg, more than 90 mg, more than 80 mg, more than 70 mg, morethan 60 mg, more than 50 mg, more than 40 mg, more than 30 mg, more than20 mg, more than 10 mg, more than 5 mg, more than 2 mg, or more than 1mg of the antiviral agent. In certain embodiments, the unit dosecomprises more than 50 mg and less than 1500 mg of the antiviral agent.In some embodiments, the unit dose comprises more than 100 mg and lessthan 500 mg of the antiviral agent. In certain embodiments, theantiviral agent is formulated as slow release.

In some embodiments, the co-formulated HDAC inhibitor and antiviralagent are administered once a day. In certain embodiments, theco-formulated HDAC inhibitor and antiviral agent are administered twicea day. In other embodiments, the co-formulated HDAC inhibitor andantiviral agent are administered thrice a day. In some embodiments, theco-formulated HDAC inhibitor and antiviral agent are administered once aday, twice a day, or thrice a day, and a further dose of the HDACinhibitor is administered once, twice, or thrice a day. In certainembodiments, the co-formulated HDAC inhibitor and antiviral agent areadministered once a day, twice a day, or thrice a day, and a furtherdose of the antiviral agent is administered once, twice, or thrice aday.

In certain embodiments, one unit dose of the co-formulated HDACinhibitor and antiviral agent are administered per day. In someembodiments, two unit doses of the co-formulated HDAC inhibitor andantiviral agent are administered per day. In certain embodiments, threeunit doses of the co-formulated HDAC inhibitor and antiviral agent areadministered per day. In some embodiments, four unit doses of theco-formulated HDAC inhibitor and antiviral agent are administered perday. In certain embodiments, the one, two, three, or four unit doses areadministered daily, once a week, twice a week, three times a week, fourtimes a week, or five times a week.

In some embodiments, one or more unit doses of the co-formulated HDACinhibitor and antiviral agent are administered in combination with othertreatments, such as antibodies, chemotherapy drugs, and radiationtherapy.

One or more agents and/or combinations of agents can be administeredwith still other agents. The choice of agents that can beco-administered with the agents and/or combinations of agents candepend, at least in part, on the condition being treated. Agents ofparticular use in the formulations of the present invention include, forexample, any agent having a therapeutic effect for a virus-inducedinflammatory condition, including, e.g., drugs used to treatinflammatory conditions. For example, formulations of the instantinvention can additionally contain one or more conventionalanti-inflammatory drugs, such as an NSAID, e.g. ibuprofen, naproxen,acetominophen, ketoprofen, or aspirin. In some alternative embodimentsfor the treatment of a virus-induced inflammatory condition canadditionally contain one or more conventional influenza antiviralagents, such as amantadine, rimantadine, zanamivir, and oseltamivir. Intreatments for retroviral infections, such as HIV, formulations of theinstant invention may additionally contain one or more conventionalantiviral drug, such as protease inhibitors (lopinavir/ritonavir{Kaletra™}, indinavir {Crixivan™}, ritonavir {Norvir™}, nelfinavir{Viracept™}, saquinavir hard gel capsules {Invirase™}, atazanavir{Reyataz™}, amprenavir {Agenerase™}, fosamprenavir {Telzir™},tipranavir{Aptivus™}), reverse transcriptase inhibitors,includingnon-Nucleoside and Nucleoside/nucleotide inhibitors (AZT{zidovudine, Retrovir™}, ddI {didanosine, Videx™}, 3TC {lamivudine,Epivir™}, d4T {stavudine, Zerit™}, abacavir {Ziagen™}, FTC{emtricitabine, Emtriva™}, tenofovir {Viread™}, efavirenz {Sustiva™} andnevirapine {Viramune™}), fusion inhibitors T20 {enfuvirtide, Fuzeon™},integrase inhibitors (MK-0518 and GS-9137), and maturation inhibitors(PA-457 {Bevirimat™}). As another example, formulations can additionallycontain one or more supplements, such as vitamin C, E or otheranti-oxidants.

One or more agents (or pharmaceutically acceptable salts, esters oramides thereof) can be administered per se or in the form of apharmaceutical composition wherein the one or more active agent(s) is inan admixture or mixture with one or more pharmaceutically acceptablecarriers. A pharmaceutical composition, as used herein, can be anycomposition prepared for administration to a subject. Pharmaceuticalcompositions can be formulated in conventional manner using one or morephysiologically acceptable carriers, comprising excipients, diluents,and/or auxiliaries, e.g., that facilitate processing of the activeagents into preparations that can be administered. Proper formulationcan depend at least in part upon the route of administration chosen. Oneor more agents, or pharmaceutically acceptable salts, esters, or amidesthereof, can be delivered to a patient using a number of routes or modesof administration, including oral, buccal, topical, rectal, transdermal,transmucosal, subcutaneous, intravenous, and intramuscular applications,as well as by inhalation.

For oral administration, one or more agents can be formulated readily bycombining the one or more active agents with pharmaceutically acceptablecarriers well known in the art. Such carriers can enable the one or moreagents to be formulated as tablets, including chewable tablets, pills,dragees, capsules, lozenges, hard candy, liquids, gels, syrups,slurries, powders, suspensions, elixirs, wafers, and the like, for oralingestion by a patient to be treated. Such formulations can comprisepharmaceutically acceptable carriers including solid diluents orfillers, sterile aqueous media and various non-toxic organic solvents.Generally, the agents of the invention can be included at concentrationlevels ranging from about 0.5%, about 5%, about 10%, about 20%, or about30% to about 50%, about 60%, about 70%, about 80% or about 90% by weightof the total composition of oral dosage forms, in an amount sufficientto provide a desired unit of dosage.

Aqueous suspensions for oral use can contain one or more agents withpharmaceutically acceptable excipients, such as a suspending agent(e.g., methyl cellulose), a wetting agent (e.g., lecithin, lysolecithinand/or a long-chain fatty alcohol), as well as coloring agents,preservatives, flavoring agents, and the like.

Oils or non-aqueous solvents can be required to bring one or more agentsinto solution, due to, for example, the presence of large lipophilicmoieties. Alternatively, emulsions, suspensions, or other preparations,for example, liposomal preparations, can be used. With respect toliposomal preparations, any known methods for preparing liposomes fortreatment of a condition can be used. See, for example, Bangham et al.,J. Mol. Biol. 23: 238-252 (1965) and Szoka et al., Proc. Natl Acad. Sci.USA 75: 4194-4198 (1978), incorporated herein by reference. Ligands canalso be attached to the liposomes to direct these compositions toparticular sites of action. One or more agents can also be integratedinto foodstuffs, e.g, cream cheese, butter, salad dressing, or ice creamto facilitate solubilization, administration, and/or compliance incertain patient populations.

Pharmaceutical preparations for oral use can be obtained as a solidexcipient, optionally grinding a resulting mixture, and processing themixture of granules, after adding suitable auxiliaries, if desired, toobtain tablets or dragee cores. Suitable excipients are, in particular,fillers such as sugars, including lactose, sucrose, mannitol, orsorbitol; flavoring elements, cellulose preparations such as, forexample, maize starch, wheat starch, rice starch, potato starch,gelatin, gum tragacanth, methyl cellulose,hydroxypropylmethyl-cellulose, sodium carboxymethylcellulose, and/orpolyvinyl pyrrolidone (PVP). Disintegrating agents can be added, forexample, the cross-linked polyvinyl pyrrolidone, agar, or alginic acidor a salt thereof such as sodium alginate. One or more agents can alsobe formulated as a sustained release preparation.

Dragee cores can be provided with suitable coatings. For this purpose,concentrated sugar solutions may be used, which may optionally containgum arabic, talc, polyvinyl pyrrolidone, carbopol gel, polyethyleneglycol, and/or titanium dioxide, lacquer solutions, and suitable organicsolvents or solvent mixtures. Dyestuffs or pigments can be added to thetablets or dragee coatings for identification or to characterizedifferent combinations of one or more active agents.

Pharmaceutical preparations that can be used orally include push-fitcapsules made of gelatin, as well as soft, sealed capsules made ofgelatin and a plasticizer, such as glycerol or sorbitol. The push-fitcapsules can contain the active ingredients in admixture with fillersuch as lactose, binders such as starches, and/or lubricants such astalc or magnesium stearate and, optionally, stabilizers. In softcapsules, the active agents can be dissolved or suspended in suitableliquids, such as fatty oils, liquid paraffin, or liquid polyethyleneglycols. In addition, stabilizers can be added. All formulations fororal administration can be in dosages suitable for administration.

For injection, one or more agents can be formulated in aqueoussolutions, including but not limited to physiologically compatiblebuffers such as Hank's solution, Ringer's solution, or physiologicalsaline buffer. Such compositions can also include one or moreexcipients, for example, preservatives, solubilizers, fillers,lubricants, stabilizers, albumin, and the like. Methods of formulationare known in the art, for example, as disclosed in Remington'sPharmaceutical Sciences, latest edition, Mack Publishing Co., Easton P.

One or more agents can also be formulated as a depot preparation. Suchlong acting formulations can be administered by implantation ortranscutaneous delivery (for example subcutaneously or intramuscularly),intramuscular injection or use of a transdermal patch. Thus, forexample, one or more agents can be formulated with suitable polymeric orhydrophobic materials (for example as an emulsion in an acceptable oil)or ion exchange resins, or as sparingly soluble derivatives, forexample, as a sparingly soluble salt.

Pharmaceutical compositions comprising one or more agents can exertlocal and regional effects when administered topically or injected at ornear particular sites of infection. Direct topical application, e.g., ofa viscous liquid, gel, jelly, cream, lotion, ointment, suppository,foam, or aerosol spray, can be used for local administration, toproduce, for example local and/or regional effects. Pharmaceuticallyappropriate vehicles for such formulation include, for example, loweraliphatic alcohols, polyglycols (e.g., glycerol or polyethylene glycol),esters of fatty acids, oils, fats, silicones, and the like. Suchpreparations may also include preservatives (e.g., p-hydroxybenzoic acidesters) and/or antioxidants (e.g., ascorbic acid and tocopherol). Seealso Dermatological Formulations: Percutaneous absorption, Barry (Ed.),Marcel Dekker Incl, 1983. In some embodiments, local/topicalformulations comprising a viral inducing agent and or antiviral agentare used to treat epidermal or mucosal viral-induced inflammatorycondition.

Pharmaceutical compositions can contain a cosmetically ordermatologically acceptable carrier. Such carriers can be compatiblewith skin, nails, mucous membranes, tissues and/or hair, and can includeany conventionally used cosmetic or dermatological carrier meeting theserequirements. Such carriers can be readily selected by one of ordinaryskill in the art. In formulating skin ointments, an agent or combinationof agents can be formulated in an oleaginous hydrocarbon base, ananhydrous absorption base, a water-in-oil absorption base, anoil-in-water water-removable base and/or a water-soluble base.

The compositions according to the present invention can be in any formsuitable for topical application, including aqueous, aqueous-alcoholicor oily solutions, lotion or serum dispersions, aqueous, anhydrous oroily gels, emulsions obtained by dispersion of a fatty phase in anaqueous phase (O/W or oil in water) or, conversely, (W/O or water inoil), microemulsions or alternatively microcapsules, microparticles orlipid vesicle dispersions of ionic and/or nonionic type. Thesecompositions can be prepared according to conventional methods. Theamounts of the various constituents of the compositions according to theinvention can be those conventionally used in the art. Thesecompositions constitute protection, treatment or care creams, milks,lotions, gels or foams for the face, for the hands, for the body and/orfor the mucous membranes, or for cleansing the skin. The compositionscan also consist of solid preparations constituting soaps or cleansingbars.

A pharmaceutical composition can also contain adjuvants common to thecosmetic and dermatological fields, for example, hydrophilic orlipophilic gelling agents, hydrophilic or lipophilic active agents,preserving agents, antioxidants, solvents, fragrances, fillers,sunscreens, odor-absorbers and dyestuffs. The amounts of these variousadjuvants can be those conventionally used in the fields considered and,for example, are from about 0.01% to about 20% of the total weight ofthe composition. Depending on their nature, these adjuvants can beintroduced into the fatty phase, into the aqueous phase and/or into thelipid vesicles.

Ocular viral infections can be effectively treated with ophthalmicsolutions, suspensions, ointments or inserts comprising an agent orcombination of agents of the present invention.

In some embodiments, viral infections of the ear can be effectivelytreated with otic solutions, suspensions, ointments or insertscomprising an agent or combination of agents of the present invention.

One or more agents can be delivered in soluble rather than suspensionform, which can allow for more rapid and quantitative absorption to thesites of action. In general, formulations such as jellies, creams,lotions, suppositories and ointments can provide an area with moreextended exposure to the agents of the present invention, whileformulations in solution, e.g., sprays, provide more immediate,short-term exposure.

Relating to topical/local application, a pharmaceutical composition caninclude one or more penetration enhancers. For example, the formulationscan comprise suitable solid or gel phase carriers or excipients thatincrease penetration or help delivery of agents or combinations ofagents of the invention across a permeability barrier, e.g., the skin.Many of these penetration-enhancing compounds are known in the art oftopical formulation, and include, e.g., water, alcohols (e.g., terpeneslike methanol, ethanol, 2-propanol), sulfoxides (e.g., dimethylsulfoxide, decylmethyl sulfoxide, tetradecylmethyl sulfoxide),pyrrolidones (e.g., 2-pyrrolidone, N-methyl-2-pyrrolidone,N-(2-hydroxyethyl)pyrrolidone), laurocapram, acetone, dimethylacetamide,dimethylformamide, tetrahydrofurfuryl alcohol, L-α-amino acids, anionic,cationic, amphoteric or nonionic surfactants (e.g., isopropyl myristateand sodium lauryl sulfate), fatty acids, fatty alcohols (e.g., oleicacid), amines, amides, clofibric acid amides, hexamethylene lauramide,proteolytic enzymes, α-bisabolol, d-limonene, urea andN,N-diethyl-m-toluamide, and the like. Additional examples includehumectants (e.g., urea), glycols (e.g., propylene glycol andpolyethylene glycol), glycerol monolaurate, alkanes, alkanols, ORGELASE,calcium carbonate, calcium phosphate, various sugars, starches,cellulose derivatives, gelatin, and/or other polymers. A pharmaceuticalcomposition can include one or more such penetration enhancers.

A pharmaceutical composition for local/topical application can includeone or more antimicrobial preservatives, for example, quaternaryammonium compounds, organic mercurials, p-hydroxy benzoates, aromaticalcohols, chlorobutanol, and the like.

Gastrointestinal viral infections can be effectively treated withorally- or rectally delivered solutions, suspensions, ointments, enemasand/or suppositories comprising an agent or combination of agents of thepresent invention.

Respiratory viral infections can be effectively treated with aerosolsolutions, suspensions or dry powders comprising an agent or combinationof agents of the present invention. Administration by inhalation isparticularly useful in treating viral infections of the lung, such asinfluenza. The aerosol can be administered through the respiratorysystem or nasal passages. For example, one skilled in the art willrecognize that a composition of the present invention can be suspendedor dissolved in an appropriate carrier, e.g., a pharmaceuticallyacceptable propellant, and administered directly into the lungs using anasal spray or inhalant. For example, an aerosol formulation comprisinga viral inducing agent and/or antiviral agent can be dissolved,suspended or emulsified in a propellant or a mixture of solvent andpropellant, e.g., for administration as a nasal spray or inhalant.Aerosol formulations may contain any acceptable propellant underpressure, such as a cosmetically or dermatologically or pharmaceuticallyacceptable propellant, as conventionally used in the art.

An aerosol formulation for nasal administration is generally an aqueoussolution designed to be administered to the nasal passages in drops orsprays. Nasal solutions can be similar to nasal secretions in that theyare generally isotonic and slightly buffered to maintain a pH of about5.5 to about 6.5, although pH values outside of this range canadditionally be used. Antimicrobial agents or preservatives can also beincluded in the formulation.

An aerosol formulation for inhalations and inhalants can be designed sothat an agent or combination of agents can be carried into therespiratory tree of the subject when administered by the nasal or oralrespiratory route. Inhalation solutions can be administered, forexample, by a nebulizer. Inhalations or insufflations, comprising finelypowdered or liquid drugs, can be delivered to the respiratory system asa pharmaceutical aerosol of a solution or suspension of the agent orcombination of agents in a propellant, e.g., to aid in disbursement.Propellants can be liquefied gases, including halocarbons, for example,fluorocarbons such as fluorinated chlorinated hydrocarbons,hydrochlorofluorocarbons, and hydrochlorocarbons, as well ashydrocarbons and hydrocarbon ethers.

Halocarbon propellants can include fluorocarbon propellants in which allhydrogens are replaced with fluorine, chlorofluorocarbon propellants inwhich all hydrogens are replaced with chlorine and at least onefluorine, hydrogen-containing fluorocarbon propellants, andhydrogen-containing chlorofluorocarbon propellants. Halocarbonpropellants are described in Johnson, U.S. Pat. No. 5,376,359, issuedDec. 27, 1994; Byron et al., U.S. Pat. No. 5,190,029, issued Mar. 2,1993; and Purewal et al., U.S. Pat. No. 5,776,434, issued Jul. 7, 1998.Hydrocarbon propellants useful in the invention include, for example,propane, isobutane, n-butane, pentane, isopentane and neopentane. Ablend of hydrocarbons can also be used as a propellant. Etherpropellants include, for example, dimethyl ether as well as the ethers.An aerosol formulation of the invention can also comprise more than onepropellant. For example, an aerosol formulation can comprise more thanone propellant from the same class, such as two or more fluorocarbons;or more than one, more than two, more than three propellants fromdifferent classes, such as a fluorohydrocarbon and a hydrocarbon.Pharmaceutical compositions of the present invention can also bedispensed with a compressed gas, e.g., an inert gas such as carbondioxide, nitrous oxide or nitrogen.

Aerosol formulations can also include other components, for example,ethanol, isopropanol, propylene glycol, as well as surfactants or othercomponents such as oils and detergents. These components can serve tostabilize the formulation and/or lubricate valve components.

The aerosol formulation can be packaged under pressure and can beformulated as an aerosol using solutions, suspensions, emulsions,powders and semisolid preparations. For example, a solution aerosolformulation can comprise a solution of an agent, such as a viralinducing agent and/or antiviral agent in (substantially) pure propellantor as a mixture of propellant and solvent. The solvent can be used todissolve the agent and/or retard the evaporation of the propellant.Solvents useful in the invention include, for example, water, ethanoland glycols. Any combination of suitable solvents can be used,optionally combined with preservatives, antioxidants, and/or otheraerosol components.

An aerosol formulation can also be a dispersion or suspension. Asuspension aerosol formulation may comprise a suspension of an agent orcombination of agents of the instant invention, e.g., a viral inducingagent and/or antiviral agent, and a dispersing agent. Dispersing agentsuseful in the invention include, for example, sorbitan trioleate, oleylalcohol, oleic acid, lecithin and corn oil. A suspension aerosolformulation can also include lubricants, preservatives, antioxidant,and/or other aerosol components.

An aerosol formulation can be formulated as an emulsion. An emulsionaerosol formulation can include, for example, an alcohol such asethanol, a surfactant, water and a propellant, as well as an agent orcombination of agents, e.g., a viral inducing agent and/or an antiviralagent. The surfactant used can be nonionic, anionic or cationic. Oneexample of an emulsion aerosol formulation comprises, for example,ethanol, surfactant, water and propellant. Another example of anemulsion aerosol formulation comprises, for example, vegetable oil,glyceryl monostearate and propane.

Pharmaceutical compositions suitable for use in the present inventioncan include compositions wherein the active ingredients are present inan effective amount, i.e., in an amount effective to achieve therapeuticand/or prophylactic benefit in a host with at least one virus-inducedinflammatory condition. The actual amount effective for a particularapplication will depend on the condition or conditions being treated,the condition of the subject, the formulation, and the route ofadministration, as well as other factors known to those of skill in theart. Determination of an effective amount of a viral inducing agentand/or antiviral agent is well within the capabilities of those skilledin the art, in light of the disclosure herein, and can be determinedusing routine optimization techniques.

An effective amount for use in humans can be determined from animalmodels. For example, a dose for humans can be formulated to achievecirculating, liver, topical and/or gastrointestinal concentrations thathave been found to be effective in animals. One skilled in the art candetermine the effective amount for human use, especially in light of theanimal model experimental data described herein. Based on animal data,and other types of similar data, those skilled in the art can determinean effective amount of a composition appropriate for humans.

An effective amount when referring to an agent or combination of agentsof the invention can generally mean the dose ranges, modes ofadministration, formulations, etc., that have been recommended orapproved by any of the various regulatory or advisory organizations inthe medical or pharmaceutical arts (e.g., FDA, AMA) or by themanufacturer or supplier.

Further, appropriate doses for a viral inducing agent and/or antiviralagent can be determined based on in vitro experimental results.

A person of skill in the art would be able to monitor in a patient theeffect of administration of a particular agent. For example, HIV or EBVviral load levels can be determined by techniques standard in the art,such as measuring CD4 cell counts, and/or viral levels as detected byPCR. Other techniques would be apparent to one of skill in the art.

Administration Schedule

Administration of one or more agents (e.g, a viral inducing agent and/oran antiviral) can be intermittent; for example, administration can beonce every two days, every three days, every five days, once a week,once or twice a month, and the like. The amount, forms, and/or amountsof the different forms can be varied at different times ofadministration.

Pulsed administration of one or more pharmaceutical compositions can beused for the treatment or prevention of a viral-induced inflammatorycondition. Pulsed administration can be more effective than continuoustreatment as pulsed doses can be lower than would be expected fromcontinuous administration of the same composition. Each pulse dose canbe reduced and the total amount of drug administered over the course oftreatment to the patient can be minimized.

With pulse therapy, in vivo levels of an agent can drop below that levelrequired for effective continuous treatment. Pulsed administration canreduce the amount of the composition administered to the patient perdose or per total treatment regimen with an increased effectiveness.Pulsed administration can provide a saving in time, effort and expenseand a lower effective dose can lessen the number and severity ofcomplications that can be experienced by a subject. As such, pulsing canbe more effective than continuous administration of the samecomposition.

Individual pulses can be delivered to a subject continuously over aperiod of several hours, such as about 2, 4, 6, 8, 10, 12, 14 or 16hours, or several days, such as 2, 3, 4, 5, 6, or 7 days, or from about1 hour to about 24 hours or from about 3 hours to about 9 hours.Alternatively, periodic doses can be administered in a single bolus or asmall number of injections of the composition over a short period oftime, for example, less than 1 or 2 hours. For example, argininebutyrate can be administered over a period of 4 days with infusions forabout 8 hours per day or overnight, followed by a period of 7 days of notreatment.

The interval between pulses or the interval of no delivery can begreater than 24 hours or can be greater than 48 hours, and can be foreven longer such as for 3, 4, 5, 6, 7, 8, 9 or 10 days, two, three orfour weeks or even longer. The interval between pulses can be determinedby one of ordinary skill in the art. The interval between pulses can becalculated by administering another dose of the composition when thecomposition or the active component of the composition is no longerdetectable in the patient prior to delivery of the next pulse. Intervalscan also be calculated from the in vivo half-life of the composition.Intervals can be calculated as greater than the in vivo half-life, or 2,3, 4, 5 and even 10 times greater than the composition half-life.Intervals can be 25, 50, 100, 150, 200, 250 300 and even 500 times thehalf life of the chemical composition.

The number of pulses in a single therapeutic regimen can be as little astwo, but can be from about 5 to 10, 10 to 20, 15 to 30 or more. Subjects(e.g., patients) can receive one or more agents (e.g., drugs) for lifeaccording to the methods of this invention. Compositions can beadministered by most any means, and can be delivered to the patient asan injection (e.g. intravenous, subcutaneous, intraarterial), infusionor instillation, and more preferably by oral ingestion. Various methodsand apparatus for pulsing compositions by infusion or other forms ofdelivery to the patient are disclosed in U.S. Pat. Nos. 4,747,825;4,723,958; 4,948,592; 4,965,251 and 5,403,590.

In certain embodiments, the co-formulated unit dose comprising an HDACinhibitor and an antiviral agent is administered daily. In furtherembodiments, administration is continuous. In some embodiments, theadministration of the co-formulated unit dose is by pulsedadministration. In certain embodiments, pulsed administration comprisesadministering pulses of the co-formulated unit dose for about 1 day,about 2 days, about 3 days, about 4 days, about 5 days, about 6 days,about 7 days, about 10 days, about 2 weeks, about 3 weeks, about 4weeks, about 5 weeks, about 6 weeks, about 7 weeks, about 8 weeks, about2 months, about 3 months, about 4 months, about 5 months, about 6months, about 9 months, about 12 months. In some embodiments, pulsedadministration comprises intervals of not administering theco-formulated unit dose of about 1 day, about 2 days, about 3 days,about 4 days, about 5 days, about 6 days, about 7 days, about 10 days,about 2 weeks, about 3 weeks, about 4 weeks, about 5 weeks, about 6weeks, about 7 weeks, about 8 weeks, about 2 months, about 3 months,about 4 months, about 5 months, about 6 months, about 9 months, about 12months.

In some embodiments, the administration of the co-formulated unit doseis by pulsed administration. In certain embodiments, the pulsedadministration comprises administering the co-formulated unit dose forabout 8 weeks, followed by not administering the co-formulated unit dosefor about 4 weeks. In some embodiments, the pulsed administrationcomprises administering the co-formulated unit dose for about 6 weeks,followed by not administering the co-formulated unit dose for about 2weeks. In certain embodiments, the pulsed administration comprisesadministering the co-formulated unit dose for about 4 weeks, followed bynot administering the co-formulated unit dose for about 2 weeks. In someembodiments, the pulsed administration comprises administering theco-formulated unit dose for about 2 weeks, followed by not administeringthe co-formulated unit dose for about 2 weeks. In some embodiments,pulsed administration comprises pulses of administering theco-formulated unit dose for about 1 day, about 2 days, about 3 days,about 4 days, about 5 days, about 6 days, about 7 days, about 10 days,about 2 weeks, about 3 weeks, about 4 weeks, about 5 weeks, about 6weeks, about 7 weeks, about 8 weeks, about 2 months, about 3 months,about 4 months, about 5 months, about 6 months, about 9 months, about 12months. In certain embodiments, pulsed administration comprisesintervals of not administering the co-formulated unit dose of about 1day, about 2 days, about 3 days, about 4 days, about 5 days, about 6days, about 7 days, about 10 days, about 2 weeks, about 3 weeks, about 4weeks, about 5 weeks, about 6 weeks, about 7 weeks, about 8 weeks, about2 months, about 3 months, about 4 months, about 5 months, about 6months, about 9 months, about 12 months. In some embodiments,administration is continuous. In certain embodiments, administration isfor the lifetime of the subject. In other embodiments, administration isfor about 1 week, about 2 weeks, about 3 weeks, about 4 weeks, about 5weeks, about 6 weeks, about 7 weeks, about 8 weeks, about 2 months,about 3 months, about 4 months, about 5 months, about 6 months, about 9months, or about 12 months. In some embodiments, an antiviral agent isadministered during intervals of not administering the co-formulatedunit dose. In certain embodiments, an antiviral agent is administered inaddition to the co-formulated unit dose. In some embodiments, anantiviral agent is administered simultaneously with the co-formulatedunit dose. In other embodiments, an antiviral agent is administeredseparate from the co-formulated unit dose.

A pharmaceutical composition comprising a viral inducing agent can beadministered to a subject before a pharmaceutical composition comprisingan antiviral agent is administered to the subject. A pharmaceuticalcomposition comprising a viral inducing agent can be co-administered toa subject with a pharmaceutical composition comprising an antiviralagent. A pharmaceutical composition comprising a viral inducing agentcan be co-administered with a pharmaceutical composition comprising anantiviral agent and a pharmaceutical composition comprising one or moreaddition agents. The pharmaceutical compositions can be provided bypulsed administration. For example, a pharmaceutical compositioncomprising a viral inducing agent can be administered to a subject,followed by administration of a pharmaceutical composition comprising anantiviral agent to the subject after an interval of time has passed, andthis order of administration the same or similar time interval can berepeated, for example, at least 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 or moretimes.

EXAMPLES Example 1. Phase 1 Study of AB Plus Ganciclovir in Patientswith EBV Associated Lymphoid Malignancies

Fifteen patients with EBV-associated lymphoid malignancies, who hadhistologically confirmed lymphoid neoplasms that were EBV+, were treatedwith AB and GCV. Prior therapies (varying in different subjects)included rituximab, chemotherapy, chemoradiotherapy and bone marrowtransplant. GCV was administered at a rate of 5 mg/kg intravenously (IV)over 1 hour twice per day, and continued throughout the cycle. AB wascontinuously infused at a starting dose of 500 mg/kg/day. Doseescalation was continued as follows until MTD was established:

Level 1: 500 mg/kg/day IV for 2 days

Level 2: 1000 mg/kg/day IV for 2 days

Level 3: 1500 mg/kg/day IV for 2 days

Level 4: 2000 mg/kg/day IV until day 21

A total of 15 patients were evaluated for anti-tumor response (Table 1).A complete response (CR) was defined as disappearance of detectablemalignant disease on imaging or physical examination (e.g., for skinlesions or tonsillar masses). A partial response (PR) was defined as a50% decrease in tumor size (the sum of the product of the largestperpendicular diameters) or measurable lesions chosen for analysis priorto beginning of treatment. For lesions which could only be measured in 1dimension, such as skin (cutaneous T cell lymphoma), a greater than 50%decrease in the largest dimension qualified as a PR. For the 3 patientswho died from co-morbidities, anti-tumor responses were confirmedpathologically at autopsy.

TABLE 1 Treatment Courses in Patients with EBV Associated LymphoidMalignancies Treated with AB and GCV Patient HD/HTD¹ Outcome, Number No.Cycles (mg/kg/day) 1 cycle Adverse Events 1 <1, 15 d  500 CR confusion;diarrhea; emesis; rejection of lung transplant* 2 <1, 16 d 1800 CRconfusion; mucositis; headache; nausea; vomiting; abdominal pain 3 <1,19 d 2000 PR confusion; mucositis; headache; nausea/vomiting; tumorregression preceding bowel perforation* 4 1 2000 PR confusion:nausea/vomiting; anorexia 5 1 2000 NR confusion; restlessness;somnolence; nausea; vomiting; abdominal pain; vision change; orthostasis6 1 1000/1000 NR Headache; nausea/vomiting; abdominal pain;thrombocytopenia 7 1 2000/1500 CR Lethargy/stupor/confusion;hypotonia/hypoesthesia; fungal infection/mucositis; tumor lysis leadingto hemorrhage* 8 1 1500/1000 PR Acoustic hallucinations; somnolence;hypokalemia; sepsis; Deep Vein Thrombosis 9 1 2000/2000 CR Confusion;fatigue; elevated BUN; tumor lysis leading to pancreatitis/hepatitis* 101 1000/800  NR Elevated BUN, encephalopathy 11 <1, 8 d  1500/1500 PRDiarrhea; hepatomegaly 12 1 2000/2000 NR Nausea; pneumonia; portinfection 13 3 938/938 PR Nausea; anorexia; weight loss; anemia;thrombocytopenia; lethargy; insomnia; hypokalemia 14 <1, 19 d 1250/1250NR Sinus, throat, back pain; thrombocytopenia; hypokalemia; lethargy 15<1, 5 d  1000/1000 PR Lethargy; increased dyspnea; polymicrobialpneumonia/acute respiratory distress syndrome *fatal AE; ¹HD/HTD—highestdose/highest tolerated dose.

Four patients were classified as achieving CRs, including 2 with PTLD, 1with extranodal NK/T cell lymphoma and 1 with peripheral T-celllymphoma. Three of these patients died after completing therapy as aresult of co-morbid conditions and complications presumed related totumor regression. Autopsy examination of 2 subjects revealed apparentcomplete disappearance of tumor, while the third patient demonstratedsignificant necrosis of residual lymphoma at autopsy.

Six patients were classified as partial responses (PRs), including 3with PTLD, 1 with diffuse large-cell B-cell lymphoma, 1 with extranodalNK/T-cell lymphoma and 1 with subcutaneous panniculitis-like T-celllymphoma.

The remaining 5 patients were classified as non-responders (NR).

In summary 10 out of 15 patients showed a degree of response totreatment of AB in combination with the antiviral ganciclovir.

Example 2. Phase II Trial of Low-Dose Arginine Butyrate andGanciclovir/Valganciclovir in EBV(+) Lymphoid Malignancies

It has previously been found that continuous infusion of inducing agent,for example, arginine butyrate, may not be necessary to maintain viralthymidine kinase expression and sensitization to anti-viral agents inEBV-associated tumors, but that, in fact, cells that survived initialexposure to the inducing agent plus the anti-viral agent remainedsusceptible to further cycles of combination treatment (Ghosh, S. K., etal. 2007 Blood Cells, Molecules, and Diseases 38:57-65, incorporatedherein in its entirety). However, it was neither anticipated norexpected that after some first period of treatment with inducing agentand anti-viral agent, one could continue the anti-viral treatmenteffectively within a cycle of therapy without continued administrationof the inducing agent (continued administration including continuedperiods of pulsing throughout).

A clinical trial was instituted utilizing a 5-day infusion of argininebutyrate and 21 days of ganciclovir/valganciclovir for EBV+ lymphomasand Post-transplant Lymphoproliferative Disorder (PTLD). The firstpatient enrolled in the protocol (with Rituximab-refractory PTLDfollowing a cord stem cell transplantation for Hodgkin Disease)tolerated the treatment regimen well, with resolution of cough withinthree days and a decrease in LDH levels.

Treatment with arginine butyrate (AB) was administered in ahospital/inpatient basis. The subject was a 32 year old with EBV-relatedpost-transplant lymphoma who had failed multiple therapies(chemotherapy, Rituxan). The subject received AB 1,000 mg/kg/doseintravenously for 5 days (day 1-5). The dose was given continuously over24 hours. AB was given through a long IV line or port due tohypertonicity. Ganciclovir at 5 mg/kg IV over 1 hour was given twice aday for five days (day 1-5). Valganciclovir 900 mg was given orallytwice per day for 16 days (day 6-21). At the end of the 21-day cycle,imaging studies were done to determine response and revealed eliminationof nearly all tumor masses (FIG. 1). Four of six target lesions resolvedcompletely, and two additional lesions decreased in size. (Table 2) Thesubject's symptoms of fever and cough resolved for first time in 9weeks. Measure of the tumor marker serum LDH was reduced from 899 to 328(normal). Additionally, EBV, CMV, and HH6 viral load becameundetectable. These findings indicate that a shorter, morepatient-accessible regimen of the virus-target therapeutic strategy ismore efficacious. Also, there remains a continuous need for an oral asopposed to an intravenous HDAC inhibitor. Therefore, the presentinvention contemplated oral HDAC inhibitors having increased potency ascompared to AB.

TABLE 2 Quantification of tumor response evaluated by CT Scan. Tumordimensions in cm. Pre-Treatment Post-Treatment Location Dimension 1Dimension 2 Dimension 1 Dimension 2 R. Upper lobe 0.7 0.7 None None R.Mid Lobe 1.1 1.1 None None R. Lower Lobe 1.4 0.8 0.8 0.6 L. Upper Lobe0.9 0.8 None None L. Lower Lobe 0.9 0.6 0.6 0.5 Lingular 0.9 0.7 NoneNone Hepatic Seg. 6 1.1 1.1 None None Hepatic Seg. 8 1.0 0.7 None NoneL. Ant. Abd. 1.1 1.9 None None Wall R. Ant. Abd. 0.9 0.5 0.9 0.5 Wall

Example 3: Analysis of Efficacy of the Herpes Anti-Virals

There are 12 mammalian HDACs, and any one of which might be required forrepression of the TK or EBV-PK gene during latency in tumors. HDACisozyme-specific siRNAs were used to knockdown individual HDACs in tumorlines expressing latent EBV to determine which one of them inducesreactivation of TK from latency, rendering it susceptible toanti-virals.

The EBV-positive B lymphoma cell line P3HR1 was used throughout theseassays. The P3HR1 cell line was originally derived from Burkitt'slymphoma patient. EBV maintains a latent state of replication in thiscell line. Cells were maintained in RPMI 1640 with 10% fetal bovineserum containing 100 U penicillin per ml and 100 μg streptomycin per ml.The HDAC inhibitors used were from five different classes: a) shortchain fatty acids, b) hydroxamic acids, c) benzamides, d) cyclictetrapeptides, and e) largazoles.

To measure the relative level of TK mRNA in various total RNApreparations, reverse transcription and quantitative PCR using real timePCR technology was used. Five micrograms of total RNA wasreverse-transcribed using random hexamer primers and Superscript IIIcDNA synthesis kit (Invitrogen). The cDNA was diluted to a final volumeof 60 μl with sterile water, 8 μl of which was then used in each realtime PCR reaction in an ABI 7500 Sequencher using SYBR-Green technology.Primers used for the amplification of TK were EBV-TK1-F:5′-AGATGACGACGGCCTCTACCA-3′ (SEQ ID NO: 1); EBV-TK1-R:5′-CCTCCTTCTGTGCACGAAGT-3′ (SEQ ID NO: 2). The ß-actin mRNA levels inthose samples were determined similarly using ß-actin-specific primersActin/hu-F: 5′-GCTCGTCGTCGACAACGGCTC-3′ (SEQ ID NO: 3); Actin/hu-R:5′-CAAACATGATCTGGGTCATCTTCTC-3′ (SEQ ID NO: 4). The relative level of TKexpression in a sample was calculated following normalization of ß-actinexpression level.

Toxicity assays with two anti-herpesvirus drugs, Gancicovir (GCV) andPenciclovir (PCV), treated to P3HR1 cells alone was conducted as acontrol. A total of 3×10⁵ P3HR1 cells were incubated with variousconcentrations of GCV or PCV and incubated for 6 days. Viable cellcounts were measured and toxicity was expressed as percentage of cellgrowth compared to untreated cells. As shown in FIG. 2A and FIG. 2B, PCVwas less toxic to the cells compared to GCV. The effect of 40 μM GCV andPCV in combination treatment approach with 1.0 mM Na-butyrate in P3HR1cells was compared (FIG. 2C). Inhibition of cell growth with 40 μM PCV(76%) was much less than with 40 μM GCV (38%). This lower level ofinhibition of cell growth with PCV did not change significantly when thedrug was used at higher concentrations (FIG. 2D).

Example 4: Analysis of Efficacy of HDAC Inhibitors

The induction of lytic phase was assayed in EBV-positive lymphoma celllines exposed to different HDAC inhibitors (HDACi) for 24-48 hrs, thenthe expression of EBV TK and other EBV transcripts by RT-PCR analysiswas quantified. To determine tumor cytotoxic activity of the combinationof HDACi and GCV, EBV+ lymphoma cells were exposed to a range ofconcentrations of HDAC inhibitors and ganciclovir (GCV) for 3 days andthen to GCV alone for another 3 days. Efficacy of a particular HDACinhibitor in the combination treatment approach was then determined byenumerating living cells. A general experimental protocol is describedbelow.

Cells

The EBV-positive B lymphoma cell line P3HR1 was used in the study. TheP3HR1 cell line was originally derived from Burkitt's lymphoma patient.EBV maintains a latent state of replication in this cell line. Thesecells were maintained in RPMI 1640 with 10% fetal bovine serumcontaining 100 U penicillin per ml and 100 μg streptomycin per ml.

Study Agent

Various HDAC inhibitors were evaluated in this study. As a positivecontrol, the Short Chain Fatty Acid butyrate, an established inducer ofEBV-TK was used. Ganciclovir (GCV) was used as the anti-viral drug.

Titration of HDAC Inhibitors on P3HR1 Cells:

Concentrations of HDAC inhibitors, which do not significantly affect theviability or proliferation of P3HR1 cells in culture were established.

Drug Sensitivity Assay

To test the sensitivity of EBV-positive lymphoma cells towards HDACinhibitors, P3HR1 cells were treated with HDAC inhibitors in thepresence of one anti-viral drug. At the end of the assay, the efficacyof the HDAC inhibitors was assessed by measuring the inhibition of cellgrowth compared to untreated cells.

Healthy, actively-growing P3HR1 cells were harvested and resuspended infresh growth media. Cells were seeded in wells. Appropriate dilutions ofHDAC inhibitors were added to certain wells, some of which received ananti-viral drug (such as GCV at 50 μM concentration). At 72 hrs, 800 μlof culture fluid was removed from each well. Wells were then refed with1.0 ml fresh growth media without HDAC inhibitors. Fresh GCV solutionwas added to the wells that originally received GCV at the same initialconcentration. On day 6, cell morphology was observed under microscopeand viable cell counts in each individual wells were determined bytrypan blue dye exclusion method using an automated cell counter(Countess, Invitrogen).

Thymidine Kinase Expression Assay

A Thymidine Kinase (TK) was used to determine if HDAC inhibitors inducedTK expression in EBV-infected lymphoma cells.

P3HR1 cells were seeded in 60 mm plates containing 3×106 cells in 3 mlof fresh growth media. Appropriate concentrations of HDAC inhibitorswere added to the plate and cells were incubated in the presence of HDACinhibitors for 6 h, 24 h, 48 h, or as needed. Cells were harvested bycentrifugation and washed once in cold PBS. Total cellular RNA was thenextracted. To measure the relative level of TK mRNA in various total RNApreparations, reverse transcription and quantitative PCR using real timePCR analysis were used. See, Ghosh, S. K., Forman, L. W., Akinsheye, I.,Perrine, S. P., Faller, D. V.: Short discontinuous sodium butyrateexposure efficiently sensitizes latently EBV infected cells towardsnucleoside analogue-mediated growth inhibition, Blood Cells Mol.Diseases (2007), 38:57-65. The relative level of TK expression in asample was calculated following normalization of ß-actin expressionlevel.

Results

The HDAC inhibitors had varying levels of synergistic activity withanti-viral agents in killing EBV+ lymphoma cells. The hydroxamic acidLBH589, the benzamide MS275, and synthetic largazole derivatives were10⁴ to 10⁵-times more potent in killing EBV+ lymphoma cells in thepresence of GCV, compared to sodium butyrate. The effectiveconcentration of LBH589 was in the range of 50-100 nM, MS275 at 200-500nM, and Largazole derivatives at 100-200 nM. At these concentrations,the drugs as single agents produced no significant growth inhibitoryactivity in the tumor cells. LBH589, MS275 and Largazole derivativesalso strongly induced EBV-TK expression in the tumor cells. In certaininstances, the effectiveness of these HDACi compounds at such lowconcentrations makes them potentially applicable as sensitizers toanti-viral therapeutics for the treatment of EBV-associated lymphomasand other viral or virally-induced conditions. In some embodiments ofthe invention, these HDAC inhibitors are used as an alternativetherapeutic option, in combination with antivirals, for the treatment ofEBV-associated tumors and other viral or virally-induced conditions.

A. Short Chain Fatty Acids

Two SCFA HDAC inhibitors, Na-butyrate (NaB) and valproic acid (VA) weretested.

Sodium Butyrate (NaB):

In a combination treatment approach, NaB+GCV reduced growth ofEBV-positive P3HR1 cells significantly (up to 50% more) compared tocells treated with NaB or GCV alone (FIG. 3A). The optimal concentrationof NaB for this purpose was found to be 1.0 mM. Responses from 1.0 mMNaB was used as a control for interpreting results in this experiment.At a higher concentration, NaB alone reduces cell growth to asignificant degree, and the synergistic effects of GCV are lost at thoseconcentrations of NaB.

Valproic Acid (VA):

The other HDACi used in this experiment, VA, also had very similaractivity (FIG. 3B). Analysis of TK mRNA level by RT and real-time PCRhowever showed that VA was less efficient than NaB in inducing TKexpression (FIG. 3C).

B. Hydroxamic Acids

A total of five different HDAC inhibitors from the hydroxamic acid groupwere examined as combination therapies. These inhibitors includescriptaid, SAHA, panobinostat-LHB589, belinostat-PXD101, and oxamflatin.All of these HDAC inhibitors can be administered orally via an oralformulation.

Scriptaid:

Scriptaid showed strong synergistic effect with GCV in reducing cellgrowth of P3HR1 cells, especially at 500 nM and 1 μM concentrations(FIG. 4A). As shown in the data, in preferred embodiments, an HDACinhibitor of the invention combined with an antiviral agent can reducethe cell count of EBV infected cells to less than 70%, 60%, 50%, 40%,30%, 20%, or 10% of cells treated without the HDAC inhibitor orantiviral treatment, or by at least 60%, 50%, 40%, 30%, or 20% fromcells treated with an antiviral agent alone.

SAHA-Vorinostat:

The combination treatment experiment with SAHA showed that it couldinduce TK expression at a higher level than that seen with efficientconcentrations of butyrate (1.0 mM) (FIG. 5B). As such, the presentinvention contemplates using an HDAC inhibitor (preferably an oral HDACinhibitor) to induce EBV kinase expression by at least 2-, 4-, 6-, 8-,or 10-fold, wherein the HDAC inhibitor is administered at aconcentration of less than 0.5 mg/kg, 1 mg/kg, 2 mg/kg, 4 mg/kg, 6mg/kg, 8 mg/kg, or 10 mg/kg.

LHB589-Panobinostat:

The growth inhibitory activity of LHB589 at a 50 nM concentration wascomparable to that of NaB at 1.0 mM (FIG. 6A). When the cells weretreated for 3 days or longer, LHB589 was extremely toxic to the cells atany concentrations 100 nM or above. Although when treated for 24 h only,cells survived well even at a concentration of 5 μM. TK expression levelin presence of LHB589 was quite high compared to optimum concentrationof NaB (2.5 mM) (FIG. 6B). Thus, in some embodiments, the presentinvention contemplates a short administration of an oral HDAC inhibitor(e.g., less than 4 days, 3 days, 2 days, 36 h, 24 h, 12 h, or 6 h) incombination with or followed by an antiviral treatment.

PXD101-Belinostat:

PXD101 induced high level of TK expression at the 5 μM concentration.(FIG. 7). Thus, the present invention contemplates using an HDACinhibitor (preferably an oral HDAC inhibitor) to induce EBV kinaseexpression by at least 5-, 10-, 15-, 20-, 25-, 30-, or 35-fold.

Oxamflatin:

Oxamflatin showed synergistic activity with GCV towards reducing cellgrowth. At a 200 nM concentration, the activity level (growthsuppression) was more than what typically seen with 1.0 mM NaB (FIG. 8).

As shown in the data, hydroxamic acid HDAC inhibitors synergisticallywith GCV reduce cell growth of P3HR1 cells. Furthermore, hydroxamic acidHDAC inhibitors induce EBV kinase expression by at least 2-, 5-, 10-,20-, or 30-fold. In addition, hydroxamic acid HDAC inhibitors incombination with antiviral agents reduce the cell count of EBV infectedcells to less than 70%, 60%, 50%, 40%, 30%, 20%, or 10% of cells treatedwithout the HDAC inhibitor or antiviral treatment.

C. Cyclic Tetrapeptide

Apicidin:

The cyclic tetrapeptide group of HDACi examined was apicidin. A toxicityassay with apicidin alone showed that concentrations of apicidin higherthan 200 nM was quite toxic to the cells. The combination treatmentassay (FIG. 9) showed that at 100 nM and 200 nM concentrations, apicidinreduced cell growth by 40-50% over cells treated with apicidin alone.However, the 200 nM concentration cell growth was significantly retardedwithout any GCV and a 500 nM concentration was very toxic to the cells.

D. Benzamide

MS-275:

Experiments show that the benzamide class of HDAC inhibitors wereextremely potent in sensitizing P3HR1 cells to GCV-mediated effects. Asshown below (FIG. 10A) a 500 nM concentration of MS-275 was as efficientas 1.0 mM NaB. Higher concentrations were extremely toxic to the cells.Interestingly, MS-275 also strongly induced TK expression at 500 nM andhigher concentrations (FIG. 10B). TK expression was also induced at only6 hr post treatment. Based on these results, an even shorter exposure toMS-275 was examined to see if it would be sufficient to sensitize P3HR1cells to GCV-mediated killing. Cells with were treated with MS-275 andGCV for shorter time periods of 24 hr or 48 hr (as opposed to 72 hr) andthen further incubated in presence of GCV for up to 6 days, at whichtime the viable cell counts were enumerated. As shown in FIG. 10C, evenat just 24 hr exposure to MS-275 sensitized the cells to GCV-mediatedeffects as efficiently as a 72 hr continuous treatment. This furtherdemonstrates that MS-275 is very effective sensitizing agent forcombination treatment studies.

E. Largazole

Largazole Derivatives:

Largazole is a member of macrocyclic depsipeptide that was originallyisolated from coral reef cyanobacteria. Largazole is a potent HDACinhibitor with specificity towards HDAC class 1 and 2 only.Additionally, largazole has very low IC 50 and HDAC isozyme specificity.16 different analogs of the largazole were tested (ab6-113b, ab6-113a,ab6-123a, ab6-123b, ab6-164b, ab6-156b, 232a, 233a, 238a, 233b, 234b,235b, 234a, 235a, 237a, 212b, TLN1 357, TNL2 380, ART01) for synergisticcell killing activity in combination with GCV (FIGS. 12A, B, C, D, andE). In some embodiments, the HDAC inhibitor and antiviral treatmentreduces the number of EBV TK induced cells by at least 10%, 20%, 30%,40%, 50%, 60%, 70%, 80%, or 90% as compared to untreated cells or cellstreated with the antiviral agent only. 13 largazole derivatives weretested both in combination treatment approach and also for their abilityto induce EBV TK (FIG. 12F). Several of the largazoles showed potentcell killing activity in combination with GCV. Thus, in someembodiments, an HDAC inhibitor of the invention is one that can induceEBV TK by at least 2-, 4-, 6-, 8-, 10-, 12-, or 14-fold as compared tocells not treated with an HDAC inhibitor. In any of the embodimentsherein, the HDAC inhibitor is preferably suitable for oral formulation.

Example 5: Analysis of Efficacy of Combination Treatment withHIV-Infected Cells

Virus production (p24 release) was examined in an HIV-1-infectedmonocyte line. Cells were treated or not treated with HDAC-inhibitorsand other compounds. P24 release expressed as optical density (“OD”)(FIG. 13), and then converted to pg of protein (FIG. 14). Argininebutyrate, phorbol myristate acetate (PMA), trichostatin A (TSA), LHB589,apicidin (API) and largazole (LARG) are shown to be active, whereas2,2-dimethyl butyrate (ST20) and 2-(quinazolin-4-ylamino)butanoic acid(RB3) increased viral production at levels similar to the control ofvehicle alone. DMSO was vehicle for some of the compounds tested.

Prophetic Example 1: Treatment of Multiple Sclerosis

To treat a subject suspected of having Epstein-Barr virus-inducedmultiple sclerosis, a health care professional administers to thesubject a dose of a pharmaceutical composition comprising a viralinducing agent, the HDAC inhibitor JNJ-26481585. A week later, a healthcare professional administers to the subject a dose of a pharmaceuticalcomposition comprising an antiviral agent, valganciclovir. A cycle ofadministration is carried out for a period of a month (FIG. 15). Duringthe period of administration of the viral inducing agent and theantiviral agent, the subject is also administered mitoxantrone to treatthe multiple sclerosis. A cycle of HDAC inhibitor and antiviraltreatment may be repeated at least 2, 4, 6, 8, 10, or 12 times asneeded.

Prophetic Example 2: Treatment of Atherosclerosis

To treat a coronary artery condition patient suspected of havingcytomegalovirus-induced atherosclerosis, a health care professionaladministers a dose of a pharmaceutical composition comprising a viralinducing agent, JNJ-26481585. A week later, a health care professionaladministers to the subject a dose of a pharmaceutical compositioncomprising valganciclovir. A cycle of administration is carried out fora period of a month. During the period of administration of the viralinducing agent and the antiviral agent, the subject is also administeredrosuvastatin to treat the atherosclerosis.

Prophetic Example 3

A patient either diagnosed with or suspected of having an Epstein BarrVirus (EBV)-associated malignancy such as nasopharyngeal carcinoma,Hodgkin's disease, Burkitt's lymphoma, post-transplantationlymphoproliferative disease, or gastric carcinoma can be treated. Ahealth care professional administers a dose of a pharmaceuticalcomposition comprising JNJ-26481585 and valganciclovir co-formulated fororal administration. The patient is administered a single daily dose intablet form, where the tablet contains 5 mg of JNJ-26481585 and 1500 mgof timed-release or slow-release valganciclovir. A cycle ofadministration is carried out for a period of 28 days. During the periodof administration, the subject can optionally also be administered anadditional chemotherapeutic agent to treat the malignancy.

While preferred embodiments of the present invention have been shown anddescribed herein, it will be obvious to those skilled in the art thatsuch embodiments are provided by way of example only. Numerousvariations, changes, and substitutions will now occur to those skilledin the art without departing from the invention. It should be understoodthat various alternatives to the embodiments of the invention describedherein may be employed in practicing the invention. It is intended thatthe following claims define the scope of the invention and that methodsand structures within the scope of these claims and their equivalents becovered thereby.

What is claimed is:
 1. A method for treating and/or preventing a viralor virally-induced condition comprising administering CHR-3996, whereinthe CHR-3996 is administered at less than 2 mg/kg per dose, and anantiviral agent, wherein the antiviral agent is selected from the groupconsisting of valganciclovir, ganciclovir, acyclovir, famciclovir, andcombinations thereof, wherein the viral or virally-induced condition iscaused by a DNA virus.
 2. The method of claim 1, wherein the DNA virusis a herpes virus.
 3. The method of claim 1, wherein the DNA virus is anEpstein-Barr virus.
 4. The method of claim 1, wherein the DNA virus is acytomegalovirus.
 5. The method of claim 1, wherein the DNA virus is avaricella zoster virus.
 6. The method of claim 1, wherein the viral orvirally-induced condition is a cancer selected from the group oflymphoma, chronic lymphocytic leukemia, nasopharyngeal carcinoma,gastric cancer, or Kaposi's sarcoma.
 7. The method of claim 1, whereinthe antiviral agent is valganciclovir.
 8. The method of claim 1, whereinthe antiviral agent is ganciclovir.
 9. The method of claim 1, whereinthe antiviral agent is acyclovir.
 10. The method of claim 1, wherein theantiviral agent is famciclovir.
 11. The method of claim 1, wherein theCHR-3996 and the antiviral agent are administered once daily.